Antibodies against immunogenic glycopeptides, compositions comprising the same and use thereof

ABSTRACT

Disclosed herein are antibodies which specifically bind to at least one epitope defined by the immunogenic glycopeptide. Other aspects of the present disclosure are pharmaceutical composition comprising the antibody; and method for preventing and/or treating Globo-H-positive cancer.

FIELD OF THE INVENTION

The present invention relates to the field of immunotherapy of cancer.More particularly, the disclosed invention relates to an antibodyagainst an immunogenic glycopeptide, a pharmaceutical composition compcomprising the antibody and to the use thereof in cancer therapy.

BACKGROUND OF THE INVENTION

Globo H is a hexasaccharide and belongs to a large number oftumor-associated carbohydrate antigens that are overexpressed on thesurface of various epithelial cancer cells, including breast, colon,ovarian, pancreatic, lung, and prostate cancer cells. The aberrantexpression of Globo H renders it an attractive candidate forimmunotherapy and the development of cancer vaccines for GloboH-positive cancers.

However, like most carbohydrate antigens, Globo H is often tolerated bythe immune system, and consequently, the immunogenicity induced by GloboH is limited. Further, the production of antibody against a specificimmunogen typically involves the cooperative interaction of two types oflymphocytes, B-cells and helper T-cells. Yet, Globo H alone cannotactivate helper T-cells, which also attributes to the poorimmunogenicity of Globo H. Accordingly, the immunization with Globo H isoften typified by low titer of immunoglobulin M (IgM) and failure toclass switch to immunoglobulin G (IgG), as well as ineffective antibodyaffinity maturation.

Various approaches have been developed to address the above-mentioneddeficiencies. In certain researches, foreign carrier proteins orpeptides having T-epitopes (such as keyhole limpet hemocyanin (KLH) ordetoxified tetanus toxoid (TT)) have been conjugated with carbohydrateantigens hoping to enhance the immunogenicity of the carbohydrateantigens. US 20010048929 provided a multivalent immunogenic molecule,comprising a carrier molecule containing at least one functional T-cellepitope, and multiple different carbohydrate fragments each linked tothe carrier molecule and each containing at least one functional B-cellepitope, wherein said carrier molecule imparts enhanced immunogenicityto said multiple carbohydrate fragments and wherein the carbohydratefragment is Globo H, LeY or STn. US 20120328646 provides a carbohydratebased vaccine containing Globo H (B cell epitope) chemically conjugatedto the immunogenic carrier diphtheria toxin cross-reacting material 197(DT-CRM 197) (Th epitope) via a p-nitrophenyl linker, which providesimmunogenicity in breast cancer models, showing delayed tumorigenesis inxenograft studies. US 20120263749 relates to a polyvalent vaccine fortreating cancer comprising at least two conjugated antigens selectedfrom a group containing glycolipid antigen such as Globo H, a Lewisantigen and a ganglioside, polysaccharide antigen, mucin antigen,glycosylated mucin antigen and an appropriate adjuvant.

Nonetheless, conjugation of carbohydrates to a carrier protein posesseveral new problems. According to Ingale et al., the foreign carrierprotein and the linker for attaching the carrier protein and thecarbohydrate may elicit strong B-cell responses, thereby leading to thesuppression of an antibody response against the carbohydrate epitope(Ingale S. et al. Robust immune responses elicited by a fully syntheticthree-component vaccine. Nat Chem Biol. 2007 October; 3(10):663-7. Epub2007 Sep. 2). Furthermore, Ingale et al. also indicated that theconjugation chemistry is difficult to control, resulting in conjugateswith ambiguities in composition and structure, which may affect thereproducibility of an immune response. Considering the above-mentionedfactors, Ingale et al. concluded that it is not surprising thatpreclinical and clinical studies using carbohydrate-protein conjugateshave led to mixed results. For example, Kuduk et al. taught that theimmunization with a trimeric cluster of Tn-antigens conjugated to KLH inthe presence of the adjuvant QS-21 elicited modest titers of IgGantibodies in mice (Kuduk S D, et al. Synthetic and immunologicalstudies on clustered modes of mucin-related Tn and TF O-linked antigens:the preparation of a glycopeptide-based vaccine for clinical trialsagainst prostate cancer. J Am Chem Soc. 1998; 120:12474-12485); whileSlovin et al. taught that the same vaccine gave low median IgG and IgMantibody titers in a clinical trial of relapsed prostate cancer patients(Slovin S F, et al. Fully synthetic carbohydrate-based vaccines inbiochemically relapsed prostate cancer: clinical trial results withalpha-N-acetylgalactosamine-O-serine/threonine conjugate vaccine. J ClinOncol. 2003; 21:4292-4298).

Moreover, for cancer patients with hypoimmune status; particular inpatients receiving chemotherapy or radiation therapy, as well aslate-stage cancer patients, the efficacy of active immune interventionis often limited, for these patients may not be able to producesufficient antibodies to elicit the anti-tumor effect.

In view of the foregoing, there exists a need in the art for developingalternative strategies for improving the immunization and/or therapeuticefficacy of carbohydrate-based vaccines.

SUMMARY OF THE INVENTION

The following presents a simplified summary of the disclosure in orderto provide a basic understanding to the reader. This summary is not anextensive overview of the disclosure and it does not identifykey/critical elements of the present invention or delineate the scope ofthe present invention. Its sole purpose is to present some conceptsdisclosed herein in a simplified form as a prelude to the more detaileddescription that is presented later.

The present disclosure is directed to an antibody which specificallybinds to Globo H according to any of the above-mentionedaspect/embodiments of the present disclosure.

According to certain embodiments, the antibody is a monoclonal antibody.

According to optional embodiments, the antibody is a chimeric orhumanized antibody.

In still another aspect, the present disclosure is directed to apharmaceutical composition for treating a cancer in a subject in needthereof.

According to one embodiment of the present disclosure, thepharmaceutical composition comprises (1) a therapeutically effectiveamount of the antibody according to any of the above-mentionedaspects/embodiments of the present disclosure, and optionally (2) apharmaceutically acceptable carrier.

In still yet another aspect, the present disclosure is directed to amethod of treating a cancer in a subject in need thereof.

According to embodiments of the present disclosure, the method includesadministering to the subject the pharmaceutical composition the antibodyor pharmaceutical composition according to any of the above-mentionedaspects/embodiments of the present disclosure.

BRIEF DESCRIPTION OF THE DRAWING

FIGS. 1A to E illustrate results of cell binding assay (A: Isotype; B:VK9; C: MZ-2; D: Control serum; E: α-Globo H serum).

FIG. 2 illustrate the binding affinity of anti-Globo H IgG antibody toprimary ovarian cancer cells according to a few working examples of thepresent disclosure.

FIG. 3 is a FACS graph illustrating the result of a serial glycan-beadbinding analysis, according to one working example of the presentdisclosure.

FIGS. 4A and B shows the simulation of protein folding of anti-Globo HIgG antibody, according to one working example of the present disclosure(A: Mouse MZ-2 monoclonal antibody; B: Humanized MZ-2 monoclonalantibody).

FIGS. 5A to E show the binding affinity of anti-Globo H IgG antibody,representing the association and Dissociation curve fitting anddissociation constant, according to one working example of the presentdisclosure (A: mMZ-2; B: cMZ-2; C:hMZ-2L; D: MK-1; E: hMZ-2Lw).

FIG. 6 shows that the human serum containing more than 1.6 μg/ml ofhMZ-2Lw antibody elicited complement-dependent cytotoxicity in breastcancer, MCF-7 cells.

FIG. 7 shows that hMZ-2Lw antibody in a concentration higher than about10 and 20 μg/ml resulted in a dose-dependent cytotoxicity in humanovarian cancer cell line TOV21G.

FIG. 8 shows that hMZ-2Lw antibody in a concentration higher than about10 and 20 μg/ml resulted in a dose-dependent cytotoxicity in humanpancreatic cancer cell line, HPAC.

FIGS. 9A and B show antigen-dependent cell-mediated cytotoxicity (ADCC)activity of the present hMZ-2Lw and MK1 antibodies on MCF-7 cells(breast cancer cell line).

FIGS. 10A and B show antigen-dependent cell-mediated cytotoxicity (ADCC)activity of the present hMZ-2Lw and MK1 antibodies on TOV21G cells(ovarian cancer cell line).

FIG. 11 shows administration of both hMZ-2Lw and MK1 antibodiessignificantly inhibit the tumor growth, while control IgG antibody didnot substantially affect the tumor growth.

FIG. 12 shows results of hMZ-2 antibody in breast cancer subcutaneousmodel MCF-7 cell.

FIGS. 13A and B shows the results of hMZ-2 antibody in pancreatic cancersubcutaneous model HPAC cell. (A: Photographs of tumors; B: Reduction oftumor size)

FIG. 14 shows that MZ-2 antibody inhibits migration of GloboH-expressing TOV21G cells.

FIG. 15 shows that MZ-2 antibody inhibits migration of GloboH-expressing TOV21G cells.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is based, at least, on the finding thatrecombinant antibodies specifically bind to Globo H for treating cancerexpression tumor-associated carbohydrate antigens.

Definition

Unless otherwise defined herein, scientific and technical terminologiesemployed in the present disclosure shall have the meanings that arecommonly understood and used by one of ordinary skill in the art. Unlessotherwise required by context, it will be understood that singular termsshall include plural forms of the same and plural terms shall includethe singular. Specifically, as used herein and in the claims, thesingular forms “a” and “an” include the plural reference unless thecontext clearly indicates otherwise. Also, as used herein and in theclaims, the terms “at least one” and “one or more” have the same meaningand include one, two, three, or more.

Notwithstanding that the numerical ranges and parameters setting forththe broad scope of the invention are approximations, the numericalvalues set forth in the specific examples are reported as precisely aspossible. Any numerical value, however, inherently contains certainerrors necessarily resulting from the standard deviation found in therespective testing measurements. Also, as used herein, the term “about”generally means within 10%, 5%, 1%, or 0.5% of a given value or range.Alternatively, the term “about” means within an acceptable standarderror of the mean when considered by one of ordinary skill in the art.Other than in the operating/working examples, or unless otherwiseexpressly specified, all of the numerical ranges, amounts, values andpercentages such as those for quantities of materials, durations oftimes, temperatures, operating conditions, ratios of amounts, and thelikes thereof disclosed herein should be understood as modified in allinstances by the term “about.” Accordingly, unless indicated to thecontrary, the numerical parameters set forth in the present disclosureand attached claims are approximations that can vary as desired. At thevery least, each numerical parameter should at least be construed inlight of the number of reported significant digits and by applyingordinary rounding techniques.

The term “antigen” as used herein is defined as a substance capable ofeliciting an immune response. Said immune response may involve eitherantibody production, or the activation of specificimmunologically-competent cells, or both. As used herein, the term“immunogen” refers to an antigen capable of inducing the production ofan antibody. Also, the term “immunogenicity” generally refers to theability of an immunogen or antigen to stimulate an immune response.

The term “epitope” refers to a unit of structure conventionally bound byan immunoglobulin VH/VL pair. An epitope defines the minimum bindingsite for an antibody, and thus represent the target of specificity of anantibody.

The term “antibody” as used herein refers to a whole antibody moleculeor a fragment, variant or derivative thereof, which is capable ofrecognizing or binding to an antigen. Most natural antibodies have twoheavy chains and two light chains linked to each other by disulfidebonds. The light chain includes a variable domain (VL) and a constantdomain (CL); while the heavy chain includes a variable domain (VH) andthree constant domains (CH1, CH2 and CH3, collectively referred to asCH). The variable regions of both light (VL) and heavy (VH) chainsdetermine binding recognition and specificity to the antigen. The VH andVL regions can be further subdivided into regions of hypervariability,termed complementarity determining regions (CDR), interspersed withregions that are more conserved, termed framework regions (FR). Each VHand VL is, composed of three CDRs and four FRs arranged fromamino-terminus to carboxy-terminus in the following order: FR1, CDR1,FR2, CDR2, FR3, CDR3, FR4. The constant region domains of the light (CL)and heavy (CH) chains confer important biological properties such asantibody chain association, secretion, trans-placental mobility,complement binding, and binding to Fc receptors (FcR).

As used herein, “antibody variable domain” refers to the portions of thelight and heavy chains of antibody molecules that include amino acidsequences of Complementarity Determining Regions (CDRs; i.e., CDR1,CDR2, and CDR3), and Framework Regions (FRs). According to the methodsused herein, the amino acid positions assigned to CDRs and FRs can bedefined according to Kabat (Sequences of Proteins of ImmunologicalInterest (National Institutes of Health, Bethesda, Md., 1987 and 1991)).Amino acid numbering of antibodies or antigen binding fragments is alsoaccording to that of Kabat.

As used herein, the term “Complementarity Determining Regions” (CDRs),i.e., CDR1, CDR2, and CDR3) refers to the amino acid residues of anantibody variable domain the presence of which are necessary for antigenbinding. Each variable domain typically has three CDR regions identifiedas CDR1, CDR2 and CDR3. H-CDR refers to the CDR of the heavy chain andL-CDR refers to the CDR of the light chain.

As used herein, the term “monoclonal antibody” refers to an antibodymolecule obtained from a single type of antibody-producing cells.

The term “chimeric antibody” refers to an antibody comprising a variableregion from one source or species and at least a portion of a constantregion derived from a different source or species, usually prepared byrecombinant DNA techniques. It is preferred that the CDRs of a chimericantibody have one origin, while the remainder of the antibody has adifferent origin. In particular, in the present invention the chimericantibody may be a humanized antibody in which the antigen bindingsequences/variable domains of a non-human antibody have been graftedonto human antibody framework regions.

As used herein, the term “humanized antibody” refers to forms ofantibodies that contain sequences from non-human (e.g., murine)antibodies as well as human antibodies. Such antibodies contain minimalsequence derived from non-human immunoglobulin. In general, thehumanized antibody will comprise substantially all of at least one, andtypically two, variable domains, in which all or substantially all ofthe hypervariable loops correspond to those of a non-humanimmunoglobulin and all or substantially all of the FR regions are thoseof a human immunoglobulin sequence. The humanized antibody optionallyalso will comprise at least a portion of an immunoglobulin constantregion (Fc), typically that of a human immunoglobulin. The humanizedforms of rodent antibodies will essentially comprise the same CDRsequences of the parental rodent antibodies, although certain amino acidsubstitutions may be included to increase affinity, increase stabilityof the humanized antibody, or for other reasons. However, as CDR loopexchanges do not uniformly result in an antibody with the same bindingproperties as the antibody of origin, changes in framework residues(FR), residues involved in CDR loop support, might also be introduced inhumanized antibodies to preserve antigen binding affinity.

Unless specified otherwise, in the peptide notation used herein, theleft-hand direction is the amino-terminal (N-terminal) direction and theright-hand direction is the carboxy-terminal (C-terminal) direction, inaccordance with standard usage and convention.

“Percentage (%) amino acid sequence identity” with respect to the aminoacid sequences identified herein is defined as the percentage of aminoacid residues in a candidate sequence that are identical with the aminoacid residues in the specific polypeptide sequence, after aligning thesequences and introducing gaps, if necessary, to achieve the maximumpercent sequence identity, and not considering any conservativesubstitutions as part of the sequence identity. Alignment for purposesof determining percentage sequence identity can be achieved in variousways that are within the skill in the art, for instance, using publiclyavailable computer software such as BLAST, BLAST-2, ALIGN or Megalign(DNASTAR) software. Those skilled in the art can determine appropriateparameters for measuring alignment, including any algorithms needed toachieve maximal alignment over the full length of the sequences beingcompared. For purposes herein, sequence comparison between two aminoacid sequences was carried out by computer program Blastp(protein-protein BLAST) provided online by Nation Center forBiotechnology Information (NCBI). Specifically, the percentage aminoacid sequence identity of a given amino acid sequence A to a given aminoacid sequence B (which can alternatively be phrased as a given aminoacid sequence A that has a certain % amino acid sequence identity to agiven amino acid sequence B) is calculated by the formula as follows:

$\frac{X}{Y} \times 100\%$

where X is the number of amino acid residues scored as identical matchesby the sequence alignment program BLAST in that program's alignment of Aand B, and where Y is the total number of amino acid residues in A or B,whichever is shorter.

As discussed herein, minor variations in the amino acid sequences ofproteins/polypeptides are contemplated as being encompassed by thepresently disclosed and claimed inventive concept(s), providing that thevariations in the amino acid sequence maintain at least 80% such as atleast, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%,94%, 95%, 96%, 97%, 98% and 99%. In particular, conservative amino acidreplacements are contemplated. Conservative replacements are those thattake place within a family of amino acids that are related in their sidechains. Genetically encoded amino acids are generally divided intofamilies: (1) acidic aspartate, glutamate; (2) basic lysine, arginine,histidine; (3) nonpolar alanine, valine, leucine, isoleucine, proline,phenylalanine, methionine, tryptophan; and (4) uncharged polar glycine,asparagine, glutamine, cysteine, serine, threonine, tyrosine. Morepreferred families are: serine and threonine are aliphatic-hydroxyfamily; asparagine and glutamine are an amide-containing family;alanine, valine, leucine and isoleucine are an aliphatic family; andphenylalanine, tryptophan, and tyrosine are an aromatic family. Forexample, it is reasonable to expect that an isolated replacement of aleucine with an isoleucine or valine, an aspartate with a glutamate, athreonine with a serine, or a similar replacement of an amino acid witha structurally related amino acid will not have a major effect on thebinding or properties of the resulting molecule, especially if thereplacement does not involve an amino acid within a framework site.Whether an amino acid change results in a functional peptide can readilybe determined by assaying the specific activity of the polypeptidederivative. Fragments or analogs of proteins/polypeptides can be readilyprepared by those of ordinary skill in the art. Preferred amino- andcarboxy-termini of fragments or analogs occur near boundaries offunctional domains.

As used herein, “antibody mutant” or “antibody variant” refers to anamino acid sequence variant of the species-dependent antibody whereinone or more of the amino acid residues of the species-dependent antibodyhave been modified. Such mutants necessarily have less than 100%sequence identity or similarity with the species-dependent antibody. Inone embodiment, the antibody mutant will have an amino acid sequencehaving at least 75% amino acid sequence identity or similarity with theamino acid sequence of either the heavy or light chain variable domainof the species-dependent antibody, more preferably at least 80%, morepreferably at least 85%, more preferably at least 90%, and mostpreferably at least 95%. Identity or similarity with respect to thissequence is defined herein as the percentage of amino acid residues inthe candidate sequence that are identical (i.e., same residue) orsimilar (i.e., amino acid residue from the same group based on commonside-chain properties, see below) with the species-dependent antibodyresidues, after aligning the sequences and introducing gaps, ifnecessary, to achieve the maximum percent sequence identity. None ofN-terminal, C-terminal, or internal extensions, deletions, or insertionsinto the antibody sequence outside of the variable domain shall beconstrued as affecting sequence identity or similarity.

As used herein, the term “antigen-binding portion” of an antibody (orsimply “antigen portion”), refers to full length or one or morefragments of an antibody that retain the ability to specifically bind toan antigen. It has been shown that the antigen-binding function of anantibody can be performed by fragments of a full-length antibody.Examples of binding fragments encompassed within the term“antigen-binding portion” of an antibody include a Fab fragment, amonovalent fragment consisting of the V_(L), V_(H), C_(L) and CH1domains; a F(ab)₂ fragment, a bivalent fragment comprising two Fabfragments linked by a disulfide bridge at the hinge region; a Fdfragment consisting of the V_(H) and CH1 domains; a Fv fragmentconsisting of the V_(L) and V_(H) domains of a single arm of anantibody; a dAb fragment (Ward et al., 1989 Nature 341:544-546), whichconsists of a V_(H) domain; and an isolated complementarity determiningregion (CDR), or any fusion proteins comprising such antigen-bindingportion.

Unless contrary to the context, the term “treatment” are used hereinbroadly to include a preventative (e.g., prophylactic), curative, orpalliative measure that results in a desired pharmaceutical and/orphysiological effect. Preferably, the effect is therapeutic in terms ofpartially or completely curing or preventing cancer. Also, the terms“treatment” and “treating” as used herein refer to application oradministration of the present immunogenic glycopeptide, antibody, orpharmaceutical composition comprising any of the above to a subject, whohas cancer, a symptom of cancer, a disease or disorder secondary tocancer, or a predisposition toward cancer, with the purpose to partiallyor completely alleviate, ameliorate, relieve, delay onset of, inhibitprogression of, reduce severity of, and/or reduce incidence of one ormore symptoms or features of cancer. Generally, a “treatment” includesnot just the improvement of symptoms or decrease of markers of thedisease, but also a cessation or slowing of progress or worsening of asymptom that would be expected in absence of treatment. The term“treating” can also be used herein in a narrower sense which refers onlyto curative or palliative measures intended to ameliorate and/or cure analready present disease state or condition in a patient or subject.

The term “preventing” as used herein refers to a preventative orprophylactic measure that stops a disease state or condition fromoccurring in a patient or subject. Prevention can also include reducingthe likelihood of a disease state or condition from occurring in apatient or subject and impeding or arresting the onset of said diseasestate or condition.

The term “effective amount” as used herein refers to the quantity of acomponent which is sufficient to yield a desired response. Effectiveamount may be expressed, for example, in grams, milligrams or microgramsor as milligrams per kilogram of body weight (mg/kg). The term alsorefers to an amount of a pharmaceutical composition containing an activecomponent or combination of components. The specific effective orsufficient amount will vary with such factors as the particularcondition being treated, the physical condition of the patient (e.g.,the patient's body mass, age, or gender), the type of mammal or animalbeing treated, the duration of the treatment, the nature of concurrenttherapy (if any), and the specific formulations employed and thestructure of the compounds or its derivatives.

As used herein, the term “therapeutically effective amount” refers tothe quantity of an active component which is sufficient to yield adesired therapeutic response. A therapeutically effective amount is alsoone in which any toxic or detrimental effects of the compound orcomposition are outweighed by the therapeutically beneficial effects.

As used herein, a “pharmaceutically acceptable carrier” is one that issuitable for use with the subjects without undue adverse side effects(such as toxicity, irritation, and allergic response) commensurate witha reasonable benefit/risk ratio. Also, each carrier must be “acceptable”in the sense of being compatible with the other ingredients of thepharmaceutical composition. The carrier can be in the form of a solid,semi-solid, or liquid diluent, cream or a capsule. The carrier must be“acceptable” in the sense of being compatible with the other ingredientsof the formulation, and is selected to minimize any degradation of theactive agent and to minimize any adverse side effects in the subject.

The term “subject” refers to a mammal including the human species thatis treatable with antibody. The term “subject” is intended to refer toboth the male and female gender unless one gender is specificallyindicated.

All patents and other publications identified are expressly incorporatedherein by reference for the purpose of describing and disclosing, forexample, the methodologies described in such publications that might beused in connection with the present invention. These publications areprovided solely for their disclosure prior to the filing date of thepresent application. Nothing in this regard should be construed as anadmission that the inventors are not entitled to antedate suchdisclosure by virtue of prior invention or for any other reason. Allstatements as to the date or representation as to the contents of thesedocuments is based on the information available to the applicants anddoes not constitute any admission as to the correctness of the dates orcontents of these documents.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as those commonly understood to one of ordinaryskill in the art to which this invention pertains. Although any knownmethods, devices, and materials may be used in the practice or testingof the invention, the methods, devices, and materials in this regard aredescribed herein.

Antibodies for Prevention and/or Treatment Cancer

Provided herein are novel recombinant anti-Globo H antibodies andanti-Globo H-binding peptides specifically binding to Globo H or itsderivatives, and methods of their use in anti-tumor immunotherapies,such as the treatment of cancer. Once bound to a cancer antigen,antibodies can induce antibody-dependent cell-mediated cytotoxicity,activate the complement system, and prevent a receptor interacting withits ligand (such as Globo H). In one embodiment, the compositionscomprising the anti-Globo H-binding peptides or anti-Globo H antibodiesdescribed herein are useful in anti-cancer therapies. Furthermore, thecompositions comprising the anti-Globo H-binding peptides or anti-GloboH antibodies can combine with an additional anti-tumor agent. Inparticular, the present embodiments provide the complementaritydetermining region (CDR) sequences of specific anti-Globo H antibodies,which can be used in a variety of anti-Globo H-binding peptides. Inparticular, the present invention provides a humanized or chimericantibody or an antigen-binding fragment thereof capable of binding toGlobo H or its derivatives.

In one aspect, the present invention provides an isolated anti-Globo Hantibody or an antigen-binding portion thereof, comprising at least oneof a heavy chain complementarity determining region 1 (H-CDR1)consisting of the amino acid residues of GFSLSTFDMGVG (SEQ ID NO: 1),GSSLSTFDVGVG (SEQ ID NO: 2), GFSLGTFDLGIG (SEQ ID NO: 3), GFSLSTFDLGIG(SEQ ID NO: 4) or a variant having amino acid sequence with at least 80%identity to any of SEQ ID NOs: 1 to 4; a heavy chain CDR2 (H-CDR2)consisting of the amino acid residues of HIWWDDDKYYNPA (SEQ ID NO:5),HIWGDDDKYYNPA (SEQ ID NO: 6) or a variant having amino acid sequencewith at least 80% identity to any of SEQ ID NOs: 5 and 6; and a heavychain CDR3 (H-CDR3) consisting of the amino acid residues ofLYGNYLTSFYCDY (SEQ ID NO: 7), or LSGNYLTSFYCDY (SEQ ID NO: 8),LYGNYLRSYYCDY (SEQ ID NO: 9) or a variant having amino acid sequencewith at least 80% identity to any of SEQ ID NOs: 7 to 9; and at leastone of a light chain CDR1 (L-CDR1) consisting of the amino acid residuesof SASSSVSYMH (SEQ ID NO: 10), SASSRVSYMH (SEQ ID NO:11), SARSSVSYMH(SEQ ID NO:12), RASSSVSYMH (SEQ ID NO:13) or a variant having amino acidsequence with at least 80% identity to any of SEQ ID NOs: 10 to 13; alight chain CDR2 (L-CDR2) consisting of the amino acid residues ofATSNLAS (SEQ ID NO:14), WTSDRYS (SEQ ID NO:15), DTSKLAS (SEQ ID NO:16)or a variant having amino acid sequence with at least 80% identity toany of SEQ ID NOs: 14 to 16; and a light chain CDR3 (L-CDR3) consistingof the amino acid residues QQWSSNPFT (SEQ ID NO: 17), QQWSSNPLT (SEQ IDNO: 18), QQHLHIPYT (SEQ ID NO: 19) or a variant having amino acidsequence with at least 80% identity to any of SEQ ID NOs: 17 to 19; suchthat said isolated antibody or antigen-binding portion thereof binds toGlobo H. Preferably, the sequence identity as mentioned above is atleast 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98% or 99%.

In a further embodiment, the present invention provides an isolatedanti-Globo H antibody or an antigen-binding portion thereof, comprising(i) a heavy chain variable region comprising a heavy chain variableregion comprising H-CDR1 selected from the group consisting of SEQ IDNOs: 1 to 4, H-CDR2 selected from the group consisting of SEQ ID NOs: 5and 6 and H-CDR3 selected from the group consisting of SEQ ID NOs: 7 to9, and (ii) light chain variable regions comprising L-CDR1 selected fromthe group consisting of SEQ ID NOs: 10 to 13, L-CDR2 selected from thegroup consisting of SEQ ID NOs: 14 to 16 and L-CDR3 selected from thegroup consisting of SEQ ID NOs: 17 to 19. Preferably, H-CDR1 is SEQ IDNO:3; H-CRD2 is SEQ ID NO:5; H-CDR3 is SEQ ID NO:8; L-CDR1 is SEQ IDNO:10; L-CDR2 is SEQ ID NO:16; and L-CDR3 is SEQ ID NO:18.

In further embodiments, H-CDR1 has the amino acid sequence of SEQ IDNO:3 or SEQ ID NO:4; H-CDR2 has the amino acid sequence of SEQ ID NO:5;H-CDR3 has the amino acid sequence of SEQ ID NO: 8; L-CDR1 has the aminoacid sequence of SEQ ID NO:12; L-CDR2 has the amino acid sequence of SEQID NO:16 and L-CDR3 has the amino acid sequence of SEQ ID NO:18.

In one aspect, the present invention provides a heavy chain variableregion comprising a heavy chain variable region comprising H-CDR1selected from the group consisting of SEQ ID NOs: 1 to 4, H-CDR2selected from the group consisting of SEQ ID NOs: 5 and 6 and H-CDR3selected from the group consisting of SEQ ID NOs: 7 to 9. In a furtherembodiment, the present invention provides a heavy chain variable regioncomprising SEQ ID NO: 3 as H-CDR1, SEQ ID NO: 5 H-CDR2 and SEQ ID NO: 8as H-CDR3.

In one aspect, the present invention provides a light chain variableregion comprising L-CDR1 selected from the group consisting of SEQ IDNOs: 10 to 13, L-CDR2 selected from the group consisting of SEQ ID NOs:14 to 16 and L-CDR3 selected from the group consisting of SEQ ID NOs: 17to 19. In a further embodiment, the present invention provides a lightchain variable region comprising SEQ ID NO: 12 as L-CDR1, SEQ ID NO: 16as L-CDR2 and SEQ ID NO: 18 as L-CDR3.

In one embodiment, the isolated anti-Globo H antibody is a monocloncalantibody. Monoclonal antibodies to Globo H can be made according toknowledge and skill in the art. For example, it can be made by injectingtest subjects with Globo H conjugate of the invention and then isolatinghybridomas expressing antibodies having the desired sequence orfunctional characteristics.

DNA encoding the monoclonal antibodies is readily isolated and sequencedusing conventional procedures (e.g., by using oligonucleotide probesthat are capable of binding specifically to genes encoding the heavy andlight chains of the monoclonal antibodies). The hybridoma cells serve asa preferred source of such DNA. Once isolated, the DNA may be placedinto expression vectors, which are then transfected into host cells suchas E. coli cells, simian COS cells, Chinese hamster ovary (CHO) cells,or myeloma cells that do not otherwise produce immunoglobulin protein,to obtain the synthesis of monoclonal antibodies in the recombinant hostcells. Recombinant production of antibodies will be described in moredetail below.

In a further embodiment, antibodies or antibody fragments can beisolated from antibody phage libraries generated using the conventionaltechniques known in the art; for example, the isolation of murine andhuman antibodies, respectively, using phage libraries. Subsequentpublications describe the production of high affinity (nM range) humanantibodies by chain shuffling, as well as combinatorial infection and invivo recombination as a strategy for constructing very large phagelibraries. Thus, these techniques are viable alternatives to traditionalmonoclonal antibody hybridoma techniques for isolation of monoclonalantibodies.

In one embodiment, the present invention provides an isolated anti-GloboH antibody, comprising (i) a heavy chain variable region comprising anamino acid sequence having at least 85% identical to any of the aminoacid sequences of SEQ ID NOs: 140 to 163, and (ii) a light chainvariable region comprising an amino acid sequence having at least 80%identical to any of the amino acid sequence of SEQ ID NO: 164 to 199.Preferably, the sequence identity as mentioned above is at least 90%,91%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%.

In one embodiment, the present invention provides an isolated anti-GloboH antibody, comprising (i) a heavy chain variable region comprising anamino acid sequence having at least 85% identical to the amino acidsequences of SEQ ID NO: 147, and (ii) a light chain variable regioncomprising an amino acid sequence having at least 80% identical to theamino acid sequence of SEQ ID NO: 195. In a further embodiment, thepresent invention provides an isolated anti-Globo H antibody, comprising(i) a heavy chain variable region comprising an amino acid sequenceconsisting of SEQ ID NO: 147, and (ii) a light chain variable regioncomprising an amino acid sequence consisting of SEQ ID NO: 195 (MZ-2antibody). The nucleotides encoding the heavy chian variable regionhaving the amino acid sequence of SEQ ID NO: 147 are as follows:

(SEQ ID NO: 236) CAGGTTACTCTGAAAGAGTCTGGCCCTGGGATATTGCAGACCTCCCAGACCCTCAGTCTGACTTGTTCTTTCTCTGGGTTTTCACTGGGCACTTTTGATTTGGGTATAGGCTGGATTCGTCAGCCTTCAGGGAAGGGTCTGGAGTGGCTGGCGCACATCTGGTGGGATGATGATAAGTACTATAATCCAGCCCTGAAGAGTCGGCTCACAATCTCCAAGGATACCTCCAAAAACCAGGTATTCCTCAAGATCGCCAATGTGGACACTGCAGACTCTGCCACATATTACTGTGCTCGGCTCTCTGGAAACTACCTCACGTCGTTCTACTGTGACTACTGGGGCCAAGGCAC CACTCTCACAGTGTCCTCAThe nucleotides encoding the light chian variable region having theamino acid sequence of SEQ ID NO: 195 are as follows:

(SEQ ID NO: 237) CAAATTGTTCTCACCCAGTCTCCAGCAATCGTGTCTGCATCTCCAGGGGAGAAGGTCACCATGACCTGCAGTGCCAGATCAAGTGTAAGTTATATGCACTGGTACCAGCAGAAGTCAGGCACCTCCCCCAAAAGATGGATTTATGACACATCCAAACTGGCTTCTGGAGTCCCTGCTCGCTTCAGTGGCAGTGGGTCTGGGACCTCTTATTCTCTCACAATCAGCAGCATGGAGGCTGAAGACGCTGCCACTTATTACTGCCAGCAGTGGAGTAGTAATCCACTCACGTTCGGTGCTGGGACCAAGCTGGAACTGAAACGG

Heavy Chain Variable Region of MZ-2 Series

(SEQ ID NO: 140) QVTLKESGPGILQTSQTLSLTCSFSGFSLSTFDMGVGWIRQPSGKGLEWLAHIWWDDDKYYNPALKSRLTISKDTSKNQVFLKIANVDTADSATYYCARL YGNYLTSFYCDYWGQGTTLTVSS (SEQ ID NO: 141)QVTLKESGPGILQTSQTLSLTCSFSGFSLSTFDMGVGWIRQPSGKGLEWLAHIWWDDDKYYNPALKSRLTISKDTSKNQVFLKIANVDTADSATYYCARL SGNYLTSFYCDYWGQGTTLTVSS (SEQ ID NO: 142)QVTLKESGPGILQTSQTLSLTCSFSGFSLSTFDMGVGWIRQPSGKGLEWLAHIWWDDDKYYNPALKSRLTISKDTSKNQVFLKIANVDTADSATYYCARLYGNYLRSYYCDYWGQGTTLTVSS (SEQ ID NO: 143)QVTLKESGPGILQTSQTLSLTCSFSGSSLSTFDVGVGWIRQPSGKGLEWLAHIWWDDDKYYNPALKSRLTISKDTSKNQVFLKIANVDTADSATYYCARLYGNYLTSFYCDYWGQGTTLTVSS (SEQ ID NO: 144)QVTLKESGPGILQTSQTLSLTCSFSGSSLSTFDVGVGWIRQPSGKGLEWLAHIWWDDDKYYNPALKSRLTISKDTSKNQVFLKIANVDTADSATYYCARLSGNYLTSFYCDYWGQGTTLTVSS (SEQ ID NO: 145)QVTLKESGPGILQTSQTLSLTCSFSGSSLSTFDVGVGWIRQPSGKGLEWLAHIWWDDDKYYNPALKSRLTISKDTSKNQVFLKIANVDTADSATYYCARLYGNYLTSFYCDYWGQGTTLTVSS (SEQ ID NO: 146)QVTLKESGPGILQTSQTLSLTCSFSGFSLGTFDLGIGWIRQPSGKGLEWLAHIWWDDDKYYNPALKSRLTISKDTSKNQVFLKIANVDTADSATYYCARLYGNYLTSFYCDYWGQGTTLTVSS (SEQ ID NO: 147)QVTLKESGPGILQTSQTLSLTCSFSGFSLGTFDLGIGWIRQPSGKGLEWLAHIWWDDDKYYNPALKSRLTISKDTSKNQVFLKIANVDTADSATYYCARLSGNYLTSFYCDYWGQGTTLTVSS (SEQ ID NO: 148)QVTLKESGPGILQTSQTLSLTCSFSGFSLGTFDLGIGWIRQPSGKGLEWLAHIWWDDDKYYNPALKSRLTISKDTSKNQVFLKIANVDTADSATYYCARLYGNYLRSYYCDYWGQGTTLTVSS  (SEQ ID NO: 149)QVTLKESGPGILQTSQTLSLTCSFSGFSLSTFDLGIGWIRQPSGKGLEWLAHIWWDDDKYYNPALKSRLTISKDTSKNQVFLKIANVDTADSATYYCARLYGNYLTSFYCDYWGQGTTLTVSS (SEQ ID NO: 150)QVTLKESGPGILQTSQTLSLTCSFSGFSLSTFDLGIGWIRQPSGKGLEWLAHIWWDDDKYYNPALKSRLTISKDTSKNQVFLKIANVDTADSATYYCARL SGNYLTSFYCDYWGQGTTLTVSS (SEQ ID NO: 151)QVTLKESGPGILQTSQTLSLTCSFSGFSLSTFDLGIGWIRQPSGKGLEWLAHIWWDDDKYYNPALKSRLTISKDTSKNQVFLKIANVDTADSATYYCARL YGNYLRSYYCDYWGQGTTLTVSS (SEQ ID NO: 152)QVTLKESGPGILQTSQTLSLTCSFSGFSLSTFDMGVGWIRQPSGKGLEWLAHIWGDDDKYYNPALKSRLTISKDTSKNQVFLKIANVDTADSATYYCARLYGNYLTSFYCDYWGQGTTLTVSS (SEQ ID NO: 153)QVTLKESGPGILQTSQTLSLTCSFSGFSLSTFDMGVGWIRQPSGKGLEWLAHIWGDDDKYYNPALKSRLTISKDTSKNQVFLKIANVDTADSATYYCARLSGNYLTSFYCDYWGQGTTLTVSS (SEQ ID NO: 154)QVTLKESGPGILQTSQTLSLTCSFSGFSLSTFDMGVGWIRQPSGKGLEWLAHIWGDDDKYYNPALKSRLTISKDTSKNQVFLKIANVDTADSATYYCARL YGNYLRSYYCDYWGQGTTLTVSS (SEQ ID NO: 155)QVTLKESGPGILQTSQTLSLTCSFSGSSLSTFDVGVGWIRQPSGKGLEWLAHIWGDDDKYYNPALKSRLTISKDTSKNQVFLKIANVDTADSATYYCARLYGNYLTSFYCDYWGQGTTLTVSS (SEQ ID NO: 156)QVTLKESGPGILQTSQTLSLTCSFSGSSLSTFDVGVGWIRQPSGKGLEWLAHIWGDDDKYYNPALKSRLTISKDTSKNQVFLKIANVDTADSATYYCARLSGNYLTSFYCDYWGQGTTLTVSS (SEQ ID NO: 157)QVTLKESGPGILQTSQTLSLTCSFSGSSLSTFDVGVGWIRQPSGKGLEWLAHIWGDDDKYYNPALKSRLTISKDTSKNQVFLKIANVDTADSATYYCARL YGNYLRSYYCDYWGQGTTLTVSS (SEQ ID NO: 158)QVTLKESGPGILQTSQTLSLTCSFSGFSLGTFDLGIGWIRQPSGKGLEWLAHIWGDDDKYYNPALKSRLTISKDTSKNQVFLKIANVDTADSATYYCARLYGNYLTSFYCDYWGQGTTLTVSS (SEQ ID NO: 159)QVTLKESGPGILQTSQTLSLTCSFSGFSLGTFDLGIGWIRQPSGKGLEWLAHIWGDDDKYYNPALKSRLTISKDTSKNQVFLKIANVDTADSATYYCARLSGNYLTSFYCDYWGQGTTLTVSS (SEQ ID NO: 160)QVTLKESGPGILQTSQTLSLTCSFSGFSLGTFDLGIGWIRQPSGKGLEWLAHIWGDDDKYYNPALKSRLTISKDTSKNQVFLKIANVDTADSATYYCARL YGNYLRSYYCDYWGQGTTLTVSS (SEQ ID NO: 161)QVTLKESGPGILQTSQTLSLTCSFSGFSLSTFDLGIGWIRQPSGKGLEWLAHIWGDDDKYYNPALKSRLTISKDTSKNQVFLKIANVDTADSATYYCARLYGNYLTSFYCDYWGQGTTLTVSS (SEQ ID NO: 162)QVTLKESGPGILQTSQTLSLTCSFSGFSLSTFDLGIGWIRQPSGKGLEWLAHIWGDDDKYYNPALKSRLTISKDTSKNQVFLKIANVDTADSATYYCARLSGNYLTSFYCDYWGQGTTLTVSS (SEQ ID NO: 163)QVTLKESGPGILQTSQTLSLTCSFSGFSLSTFDLGIGWIRQPSGKGLEWLAHIWGDDDKYYNPALKSRLTISKDTSKNQVFLKIANVDTADSATYYCARLYGNYLRSYYCDYWGQGTTLTVSS

Light Chain Variable Region of MZ-2 Series

(SEQ ID NO: 164) QIVLTQSPAIVSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYATSNLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPFTFGAG TKLELKR (SEQ ID NO: 165) QIVLTQSPAIVSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYATSNLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPLTFGAG TKLELKR(SEQ ID NO: 166) QIVLTQSPAIVSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYATSNLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQHLHIPYTFGAG TKLELKR(SEQ ID NO: 167) QIVLTQSPAIVSASPGEKVTMTCSASSRVSYMHWYQQKSGTSPKRWIYATSNLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPFTFGAG TKLELKR(SEQ ID NO: 168) QIVLTQSPAIVSASPGEKVTMTCSASSRVSYMHWYQQKSGTSPKRWIYATSNLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPLTFGAG TKLELKR(SEQ ID NO: 169) QIVLTQSPAIVSASPGEKVTMTCSASSRVSYMHWYQQKSGTSPKRWIYATSNLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQHLHIPYTFGAG TKLELKR(SEQ ID NO: 170) QIVLTQSPAIVSASPGEKVTMTCSARSSVSYMHWYQQKSGTSPKRWIYATSNLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPFTFGAG TKLELKR(SEQ ID NO: 171) QIVLTQSPAIVSASPGEKVTMTCSARSSVSYMHWYQQKSGTSPKRWIYATSNLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPLTFGAG TKLELKR(SEQ ID NO: 172) QIVLTQSPAIVSASPGEKVTMTCSARSSVSYMHWYQQKSGTSPKRWIYATSNLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQHLHIPYTFGAG TKLELKR(SEQ ID NO: 173) QIVLTQSPAIVSASPGEKVTMTCRASSSVSYMHWYQQKSGTSPKRWIYATSNLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPFTFGAG TKLELKR(SEQ ID NO: 174) QIVLTQSPAIVSASPGEKVTMTCRASSSVSYMHWYQQKSGTSPKRWIYATSNLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPLTFGAG TKLELKR(SEQ ID NO: 175) QIVLTQSPAIVSASPGEKVTMTCRASSSVSYMHWYQQKSGTSPKRWIYATSNLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQHLHIPYTFGAG TKLELKR(SEQ ID NO: 176) QIVLTQSPAIVSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYWTSDRYSGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPFTFGAG TKLELKR(SEQ ID NO: 177) QIVLTQSPAIVSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYWTSDRYSGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPLTFGAG TKLELKR(SEQ ID NO: 178) QIVLTQSPAIVSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYWTSDRYSGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQHLHIPYTFGAG TKLELKR(SEQ ID NO: 179) QIVLTQSPAIVSASPGEKVTMTCSASSRVSYMHWYQQKSGTSPKRWIYWTSDRYSGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPFTFGAG TKLELKR(SEQ ID NO: 180) QIVLTQSPAIVSASPGEKVTMTCSASSRVSYMHWYQQKSGTSPKRWIYWTSDRYSGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPLTF GAGTKLELKR(SEQ ID NO: 181) QIVLTQSPAIVSASPGEKVTMTCSASSRVSYMHWYQQKSGTSPKRWIYWTSDRYSGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQHLHIPYTFG AGTKLELKR(SEQ ID NO: 182) QIVLTQSPAIVSASPGEKVTMTCSARSSVSYMHWYQQKSGTSPKRWIYWTSDRYSGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPFTFGAG TKLELKR(SEQ ID NO: 183) QIVLTQSPAIVSASPGEKVTMTCSARSSVSYMHWYQQKSGTSPKRWIYWTSDRYSGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPLTFGAG TKLELKR(SEQ ID NO: 184) QIVLTQSPAIVSASPGEKVTMTCSARSSVSYMHWYQQKSGTSPKRWIYWTSDRYSGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQHLHIPYTFGAG TKLELKR(SEQ ID NO: 185) QIVLTQSPAIVSASPGEKVTMTCRASSSVSYMHWYQQKSGTSPKRWIYWTSDRYSGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPFTFGAG TKLELKR(SEQ ID NO: 186) QIVLTQSPAIVSASPGEKVTMTCRASSSVSYMHWYQQKSGTSPKRWIYWTSDRYSGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPLTFGAG TKLELKR(SEQ ID NO: 187) QIVLTQSPAIVSASPGEKVTMTCRASSSVSYMHWYQQKSGTSPKRWIYWTSDRYSGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQHLHIPYTFGAG TKLELKR(SEQ ID NO: 188) QIVLTQSPAIVSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPFTFGAG TKLELKR(SEQ ID NO: 189) QIVLTQSPAIVSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPLTFGAG TKLELKR(SEQ ID NO: 190) QIVLTQSPAIVSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQHLHIPYTFGAG TKLELKR(SEQ ID NO: 191) QIVLTQSPAIVSASPGEKVTMTCSASSRVSYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPFTFGAG TKLELKR(SEQ ID NO: 192) QIVLTQSPAIVSASPGEKVTMTCSASSRVSYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPLTFGAG TKLELKR(SEQ ID NO: 193) QIVLTQSPAIVSASPGEKVTMTCSASSRVSYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQHLHIPYTFGAG TKLELKR(SEQ ID NO: 194) QIVLTQSPAIVSASPGEKVTMTCSARSSVSYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPFTFGAG TKLELKR(SEQ ID NO: 195) QIVLTQSPAIVSASPGEKVTMTCSARSSVSYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPLTFGAG TKLELKR(SEQ ID NO: 196) QIVLTQSPAIVSASPGEKVTMTCSARSSVSYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQHLHIPYTFGAG TKLELKR(SEQ ID NO: 197) QIVLTQSPAIVSASPGEKVTMTCRASSSVSYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPFTFGAG TKLELKR(SEQ ID NO: 198) QIVLTQSPAIVSASPGEKVTMTCSARSSVSYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPLTFGAG TKLELKR(SEQ ID NO: 199) QIVLTQSPAIVSASPGEKVTMTCSARSSVSYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQHLHIPYTFGAG TKLELKR

In another embodiment, the isolated anti-Globo H antibody of the presentinvention is a humanized or chimeric antibody or fragment thereofcapable of specifically binding to Globo H.

In a further embodiment, the present invention provides a humanizedanti-Globo H antibody or an antigen-binding portion thereof, comprising(i) a heavy chain variable region comprising an amino acid sequencehaving at least 85% identical to any of the amino acid sequences of SEQID NOs: 20 to 43, and (ii) a light chain variable region comprising anamino acid sequence having at least 80% identical to any of the aminoacid sequence of SEQ ID NO: 44 to 79 and 200 to 235. Preferably, thesequence identity as mentioned above is at least 90%, 91%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98% or 99%.

In another further embodiment, the present invention provides ahumanized anti-Globo H antibody or an antigen-binding portion thereof,comprising (i) a heavy chain variable region comprising an amino acidsequence selected from the group consisting of SEQ ID NOs: 20 to 43, and(ii) a light chain variable region comprising an amino acid sequenceselected from the group consisting of SEQ ID NO: 44 to 79 and 200 to235. In another embodiment, the humanized anti-Globo H antibody or anantigen-binding portion thereof comprises a heavy chain variable regioncomprising an amino acid sequence consisting of SEQ ID NO: 27, and alight chain variable region comprising an amino acid sequence consistingof SEQ ID NO: 75 (hMZ-2Lw antibody). In another embodiment, thehumanized anti-Globo H antibody or an antigen-binding portion thereofcomprises a heavy chain variable region comprising an amino acidsequence consisting of SEQ ID NO: 27, and a light chain variable regioncomprising an amino acid sequence consisting of SEQ ID NO: 231.

Heavy Chain Variable Region of hMZ-2 Series

(QITLKESGPTLVKPTQTLTLTCTFS************WIRQPPGKALEWLA*************LKSRLTITKDTSKNQVVLTMTNMDPVDTATYYCAR*************WGQGTLVTVSS) (SEQ ID NO: 20)QITLKESGPTLVKPTQTLTLTCTFSGFSLSTFDMGVGWIRQPPGKALEWLAHIWWDDDKYYNPALKSRLTITKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 21)QITLKESGPTLVKPTQTLTLTCTFSGFSLSTFDMGVGWIRQPPGKALEWLAHIWWDDDKYYNPALKSRLTITKDTSKNQVVLTMTNMDPVDTATYYCARLSGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 22)QITLKESGPTLVKPTQTLTLTCTFSGFSLSTFDMGVGWIRQPPGKALEWLAHIWWDDDKYYNPALKSRLTITKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLRSYYCDYWGQGTLVTVSS (SEQ ID NO: 23)QITLKESGPTLVKPTQTLTLTCTFSGSSLSTFDVGVGWIRQPPGKALEWLAHIWWDDDKYYNPALKSRLTITKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 24)QITLKESGPTLVKPTQTLTLTCTFSGSSLSTFDVGVGWIRQPPGKALEWLAHIWWDDDKYYNPALKSRLTITKDTSKNQVVLTMTNMDPVDTATYYCARLSGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 25)QITLKESGPTLVKPTQTLTLTCTFSGSSLSTFDVGVGWIRQPPGKALEWLAHIWWDDDKYYNPALKSRLTITKDTSKNQVVLTMTNMDPVDTATYYCARLSGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 26)QITLKESGPTLVKPTQTLTLTCTFSGFSLGTFDLGIGWIRQPPGKALEWLAHIWWDDDKYYNPALKSRLTITKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 27)QITLKESGPTLVKPTQTLTLTCTFSGFSLGTFDLGIGWIRQPPGKALEWLAHIWWDDDKYYNPALKSRLTITKDTSKNQVVLTMTNMDPVDTATYYCARLSGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 28)QITLKESGPTLVKPTQTLTLTCTFSGFSLGTFDLGIGWIRQPPGKALEWLAHIWWDDDKYYNPALKSRLTITKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLRSYYCDYWGQGTLVTVSS (SEQ ID NO: 29)QITLKESGPTLVKPTQTLTLTCTFSGFSLSTFDLGIGWIRQPPGKALEWLAHIWWDDDKYYNPALKSRLTITKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 30)QITLKESGPTLVKPTQTLTLTCTFSGFSLSTFDLGIGWIRQPPGKALEWLAHIWWDDDKYYNPALKSRLTITKDTSKNQVVLTMTNMDPVDTATYYCARLSGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 31)QITLKESGPTLVKPTQTLTLTCTFSGFSLSTFDLGIGWIRQPPGKALEWLAHIWWDDDKYYNPALKSRLTITKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLRSYYCDYWGQGTLVTVSS (SEQ ID NO: 32)QITLKESGPTLVKPTQTLTLTCTFSGFSLSTFDMGVGWIRQPPGKALEWLAHIWGDDDKYYNPALKSRLTITKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 33)QITLKESGPTLVKPTQTLTLTCTFSGFSLSTFDMGVGWIRQPPGKALEWLAHIWGDDDKYYNPALKSRLTITKDTSKNQVVLTMTNMDPVDTATYYCARLSGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 34)QITLKESGPTLVKPTQTLTLTCTFSGFSLSTFDMGVGWIRQPPGKALEWLAHIWGDDDKYYNPALKSRLTITKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLRSYYCDYWGQGTLVTVSS (SEQ ID NO: 35)QITLKESGPTLVKPTQTLTLTCTFSGSSLSTFDVGVGWIRQPPGKALEWLAHIWGDDDKYYNPALKSRLTITKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 36)QITLKESGPTLVKPTQTLTLTCTFSGSSLSTFDVGVGWIRQPPGKALEWLAHIWGDDDKYYNPALKSRLTITKDTSKNQVVLTMTNMDPVDTATYYCARLSGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 37)QITLKESGPTLVKPTQTLTLTCTFSGSSLSTFDVGVGWIRQPPGKALEWLAHIWGDDDKYYNPALKSRLTITKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLRSYYCDYWGQGTLVTVSS (SEQ ID NO: 38)QITLKESGPTLVKPTQTLTLTCTFSGFSLGTFDLGIGWIRQPPGKALEWLAHIWGDDDKYYNPALKSRLTITKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 39)QITLKESGPTLVKPTQTLTLTCTFSGFSLGTFDLGIGWIRQPPGKALEWLAHIWGDDDKYYNPALKSRLTITKDTSKNQVVLTMTNMDPVDTATYYCARLSGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 40)QITLKESGPTLVKPTQTLTLTCTFSGFSLGTFDLGIGWIRQPPGKALEWLAHIWGDDDKYYNPALKSRLTITKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLRSYYCDYWGQGTLVTVSS (SEQ ID NO: 41)QITLKESGPTLVKPTQTLTLTCTFSGFSLSTFDLGIGWIRQPPGKALEWLAHIWGDDDKYYNPALKSRLTITKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 42)QITLKESGPTLVKPTQTLTLTCTFSGFSLSTFDLGIGWIRQPPGKALEWLAHIWGDDDKYYNPALKSRLTITKDTSKNQVVLTMTNMDPVDTATYYCARLSGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 43)QITLKESGPTLVKPTQTLTLTCTFSGFSLSTFDLGIGWIRQPPGKALEWLAHIWGDDDKYYNPALKSRLTITKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLRSYYCDYWGQGTLVTVSSLight Chain Variable Region of hMZ-2 Series

(EIVLTQSPSSLSASVGDRVTITC**********WYQQKPGKAPKLLIY*******GVPSRFSGSGSGTDFTFTISSLQPEDIATYYC*********FGG GTKLEIKR)(SEQ ID NO: 44)EIVLTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKPGKAPKLLIYATSNLASGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQWSSNPFTFGGGTKLEIKR (SEQ ID NO: 45)EIVLTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKPGKAPKLLIYATSNLASGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQWSSNPLTFGGGTKLEIKR (SEQ ID NO: 46)EIVLTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKPGKAPKLLIYATSNLASGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQHLHIPYTFGGGTKLEIKR (SEQ ID NO: 47)EIVLTQSPSSLSASVGDRVTITCSASSRVSYMHWYQQKPGKAPKLLIYATSNLASGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQWSSNPFTFGGGTKLEIKR (SEQ ID NO: 48)EIVLTQSPSSLSASVGDRVTITCSASSRVSYMHWYQQKPGKAPKLLIYATSNLASGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQWSSNPLTFGGGTKLEIKR (SEQ ID NO: 49)EIVLTQSPSSLSASVGDRVTITCSASSRVSYMHWYQQKPGKAPKLLIYATSNLASGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQHLHIPYTFGGGTKLEIKR (SEQ ID NO: 50)EIVLTQSPSSLSASVGDRVTITCSARSSVSYMHWYQQKPGKAPKLLIYATSNLASGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQWSSNPFTFGGGTKLEIKR (SEQ ID NO: 51)EIVLTQSPSSLSASVGDRVTITCSARSSVSYMHWYQQKPGKAPKLLIYATSNLASGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQWSSNPLTFGGGTKLEIKR (SEQ ID NO: 52)EIVLTQSPSSLSASVGDRVTITCSARSSVSYMHWYQQKPGKAPKLLIYATSNLASGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQHLHIPYTFGGGTKLEIKR (SEQ ID NO: 53)EIVLTQSPSSLSASVGDRVTITCRASSSVSYMHWYQQKPGKAPKLLIYATSNLASGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQWSSNPFTFGGGTKLEIKR (SEQ ID NO: 54)EIVLTQSPSSLSASVGDRVTITCRASSSVSYMHWYQQKPGKAPKLLIYATSNLASGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQWSSNPLTFGGGTKLEIKR (SEQ ID NO: 55)EIVLTQSPSSLSASVGDRVTITCRASSSVSYMHWYQQKPGKAPKLLIYATSNLASGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQHLHIPYTFGGGTKLEIKR (SEQ ID NO: 56)EIVLTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKPGKAPKLLIYWTSDRYSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQWSSNPFTFGGGTKLEIKR (SEQ ID NO: 57)EIVLTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKPGKAPKLLIYWTSDRYSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQWSSNPLTFGGGTKLEIKR (SEQ ID NO: 58)EIVLTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKPGKAPKLLIYWTSDRYSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQHLHIPYTFGGGTKLEIKR (SEQ ID NO: 59)EIVLTQSPSSLSASVGDRVTITCSASSRVSYMHWYQQKPGKAPKLLIYWTSDRYSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQWSSNPFTFGGGTKLEIKR (SEQ ID NO: 60)EIVLTQSPSSLSASVGDRVTITCSASSRVSYMHWYQQKPGKAPKLLIYWTSDRYSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQWSSNPLTFGGGTKLEIKR (SEQ ID NO: 61)EIVLTQSPSSLSASVGDRVTITCSASSRVSYMHWYQQKPGKAPKLLIYWTSDRYSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQHLHIPYTFGGGTKLEIKR (SEQ ID NO: 62)EIVLTQSPSSLSASVGDRVTITCSARSSVSYMHWYQQKPGKAPKLLIYWTSDRYSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQWSSNPFTFGGGTKLEIKR (SEQ ID NO: 63)EIVLTQSPSSLSASVGDRVTITCSARSSVSYMHWYQQKPGKAPKLLIYWTSDRYSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQWSSNPLTFGGGTKLEIKR (SEQ ID NO: 64)EIVLTQSPSSLSASVGDRVTITCSARSSVSYMHWYQQKPGKAPKLLIYWTSDRYSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQHLHIPYTFGGGTKLEIKR (SEQ ID NO: 65)EIVLTQSPSSLSASVGDRVTITCRASSSVSYMHWYQQKPGKAPKLLIYWTSDRYSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQWSSNPFTFGGGTKLEIKR (SEQ ID NO: 66)EIVLTQSPSSLSASVGDRVTITCRASSSVSYMHWYQQKPGKAPKLLIYWTSDRYSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQWSSNPLTFGGGTKLEIKR (SEQ ID NO: 67)EIVLTQSPSSLSASVGDRVTITCRASSSVSYMHWYQQKPGKAPKLLIYWTSDRYSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQHLHIPYTFGGGTKLEIKR (SEQ ID NO: 68)EIVLTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKPGKAPKLLIYDTSKLASGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQWSSNPFTFGGGTKLEIKR (SEQ ID NO: 69)EIVLTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKPGKAPKLLIYDTSKLASGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQWSSNPLTFGGGTKLEIKR (SEQ ID NO: 70)EIVLTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKPGKAPKLLIYDTSKLASGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQHLHIPYTFGGGTKLEIKR (SEQ ID NO: 71)EIVLTQSPSSLSASVGDRVTITCSASSRVSYMHWYQQKPGKAPKLLIYDTSKLASGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQWSSNPFTFGGGTKLEIKR (SEQ ID NO: 72)EIVLTQSPSSLSASVGDRVTITCSASSRVSYMHWYQQKPGKAPKLLIYDTSKLASGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQWSSNPLTFGGGTKLEIKR (SEQ ID NO: 73)EIVLTQSPSSLSASVGDRVTITCSASSRVSYMHWYQQKPGKAPKLLIYDTSKLASGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQHLHIPYTFGGGTKLEIKR (SEQ ID NO: 74)EIVLTQSPSSLSASVGDRVTITCSARSSVSYMHWYQQKPGKAPKLLIYDTSKLASGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQWSSNPFTFGGGTKLEIKR (SEQ ID NO: 75)EIVLTQSPSSLSASVGDRVTITCSARSSVSYMHWYQQKPGKAPKLLIYDTSKLASGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQWSSNPLTFGGGTKLEIKR (SEQ ID NO: 76)EIVLTQSPSSLSASVGDRVTITCSARSSVSYMHWYQQKPGKAPKLLIYDTSKLASGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQHLHIPYTFGGGTKLEIKR (SEQ ID NO: 77)EIVLTQSPSSLSASVGDRVTITCRASSSVSYMHWYQQKPGKAPKLLIYDTSKLASGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQWSSNPFTFGGGTKLEIKR (SEQ ID NO: 78)EIVLTQSPSSLSASVGDRVTITCRASSSVSYMHWYQQKPGKAPKLLIYDTSKLASGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQWSSNPLTFGGGTKLEIKR (SEQ ID NO: 79)EIVLTQSPSSLSASVGDRVTITCRASSSVSYMHWYQQKPGKAPKLLIYDTSKLASGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQHLHIPYTFGGGTKLEIKR (SEQ ID NO: 200)EIQMTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKPGKAPKLLIYATSNLASGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQWSSNPFTFGGGTKLEIKR (SEQ ID NO: 201)EIQMTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKPGKAPKLLIYATSNLASGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQWSSNPLTFGGGTKLEIKR (SEQ ID NO: 202)EIQMTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKPGKAPKLLIYATSNLASGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQHLHIPYTFGGGTKLEIKR (SEQ ID NO: 203)EIQMTQSPSSLSASVGDRVTITCSASSRVSYMHWYQQKPGKAPKLLIYATSNLASGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQWSSNPFTFGGGTKLEIKR (SEQ ID NO: 204)EIQMTQSPSSLSASVGDRVTITCSASSRVSYMHWYQQKPGKAPKLLIYATSNLASGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQWSSNPLTFGGGTKLEIKR (SEQ ID NO: 205)EIQMTQSPSSLSASVGDRVTITCSASSRVSYMHWYQQKPGKAPKLLIYATSNLASGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQHLHIPYTFGGGTKLEIKR (SEQ ID NO: 206)EIQMTQSPSSLSASVGDRVTITCSARSSVSYMHWYQQKPGKAPKLLIYATSNLASGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQWSSNPFTFGGGTKLEIKR (SEQ ID NO: 207)EIQMTQSPSSLSASVGDRVTITCSARSSVSYMHWYQQKPGKAPKLLIYATSNLASGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQWSSNPLTFGGGTKLEIKR (SEQ ID NO: 208)EIQMTQSPSSLSASVGDRVTITCSARSSVSYMHWYQQKPGKAPKLLIYATSNLASGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQHLHIPYTFGGGTKLEIKR (SEQ ID NO: 209)EIQMTQSPSSLSASVGDRVTITCRASSSVSYMHWYQQKPGKAPKLLIYATSNLASGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQWSSNPFTFGGGTKLEIKR (SEQ ID NO: 210)EIQMTQSPSSLSASVGDRVTITCRASSSVSYMHWYQQKPGKAPKLLIYATSNLASGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQWSSNPLTFGGGTKLEIKR (SEQ ID NO: 211)EIQMTQSPSSLSASVGDRVTITCRASSSVSYMHWYQQKPGKAPKLLIYATSNLASGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQHLHIPYTFGGGTKLEIKR (SEQ ID NO: 212)EIQMTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKPGKAPKLLIYWTSDRYSGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQWSSNPFTFGGGTKLEIKR (SEQ ID NO: 213)EIQMTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKPGKAPKLLIYWTSDRYSGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQWSSNPLTFGGGTKLEIKR (SEQ ID NO: 214)EIQMTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKPGKAPKLLIYWTSDRYSGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQHLHIPYTFGGGTKLEIKR (SEQ ID NO: 215)EIQMTQSPSSLSASVGDRVTITCSASSRVSYMHWYQQKPGKAPKWYWTSDRYSGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQWSSNPFTFGGGTKLEIKR (SEQ ID NO: 216)EIQMTQSPSSLSASVGDRVTITCSASSRVSYMHWYQQKPGKAPKWYWTSDRYSGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQWSSNPLTFGGGTKLEIKR (SEQ ID NO: 217)EIQMTQSPSSLSASVGDRVTITCSASSRVSYMHWYQQKPGKAPKWYWTSDRYSGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQHLHIPYTFGGGTKLEIKR (SEQ ID NO: 218)EIQMTQSPSSLSASVGDRVTITCSARSSVSYMHWYQQKPGKAPKWYWTSDRYSGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQWSSNPFTFGGGTKLEIKR (SEQ ID NO: 219)EIQMTQSPSSLSASVGDRVTITCSARSSVSYMHWYQQKPGKAPKWYWTSDRYSGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQWSSNPLTFGGGTKLEIKR (SEQ ID NO: 220)EIQMTQSPSSLSASVGDRVTITCSARSSVSYMHWYQQKPGKAPKWYWTSDRYSGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQHLHIPYTFGGGTKLEIKR (SEQ ID NO: 221)EIQMTQSPSSLSASVGDRVTITCRASSSVSYMHWYQQKPGKAPKWYWTSDRYSGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQWSSNPFTFGGGTKLEIKR (SEQ ID NO: 222)EIQMTQSPSSLSASVGDRVTITCRASSSVSYMHWYQQKPGKAPKWYWTSDRYSGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQWSSNPLTFGGGTKLEIKR (SEQ ID NO: 223)EIQMTQSPSSLSASVGDRVTITCRASSSVSYMHWYQQKPGKAPKWYWTSDRYSGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQHLHIPYTFGGGTKLEIKR (SEQ ID NO: 224)EIQMTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKPGKAPKLLIYDTSKLASGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQWSSNPFTFGGGTKLEIKR (SEQ ID NO: 225)EIQMTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKPGKAPKLLIYDTSKLASGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQWSSNPLTFGGGTKLEIKR (SEQ ID NO: 226)EIQMTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKPGKAPKLLIYDTSKLASGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQHLHIPYTFGGGTKLEIKR (SEQ ID NO: 227)EIQMTQSPSSLSASVGDRVTITCSASSRVSYMHWYQQKPGKAPKWYDTSKLASGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQWSSNPFTFGGGTKLEIKR (SEQ ID NO: 228)EIQMTQSPSSLSASVGDRVTITCSASSRVSYMHWYQQKPGKAPKWYDTSKLASGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQWSSNPLTFGGGTKLEIKR (SEQ ID NO: 229)EIQMTQSPSSLSASVGDRVTITCSASSRVSYMHWYQQKPGKAPKWYDTSKLASGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQHLHIPYTFGGGTKLEIKR (SEQ ID NO: 230)EIQMTQSPSSLSASVGDRVTITCSARSSVSYMHWYQQKPGKAPKWYDTSKLASGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQWSSNPFTFGGGTKLEIKR (SEQ ID NO: 231)EIQMTQSPSSLSASVGDRVTITCSARSSVSYMHWYQQKPGKAPKWYDTSKLASGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQWSSNPLTFGGGTKLEIKR (SEQ ID NO: 232)EIQMTQSPSSLSASVGDRVTITCSARSSVSYMHWYQQKPGKAPKWYDTSKLASGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQHLHIPYTFGGGTKLEIKR (SEQ ID NO: 233)EIQMTQSPSSLSASVGDRVTITCRASSSVSYMHWYQQKPGKAPKWYDTSKLASGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQWSSNPFTFGGGTKLEIKR (SEQ ID NO: 234)EIQMTQSPSSLSASVGDRVTITCRASSSVSYMHWYQQKPGKAPKWYDTSKLASGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQWSSNPLTFGGGTKLEIKR (SEQ ID NO: 235)EIQMTQSPSSLSASVGDRVTITCRASSSVSYMHWYQQKPGKAPKWYDTSKLASGVPSRFSGSGSGTDFTLTISSLQPEDIATYYCQQHLHIPYTFGGGTKLEIKR

In a further embodiment, the present invention provides a humanizedanti-Globo H antibody or an antigen-binding portion thereof, comprising(i) a heavy chain variable region comprising an amino acid sequencehaving at least 85% identical to any of the amino acid sequences of SEQID NOs: 80 to 103, and (ii) a light chain variable region comprising anamino acid sequence having at least 80% identical to any of the aminoacid sequence of SEQ ID NOs: 104 to 139. Preferably, the sequenceidentity as mentioned above is at least 90%, 91%, 91%, 92%, 93%, 94%,95%, 96%, 97%, 98% or 99%.

In another further embodiment, the present invention provides ahumanized anti-Globo H antibody or an antigen-binding portion thereof,comprising (i) a heavy chain variable region comprising an amino acidsequence selected from the group consisting of SEQ ID NOs: 80 to 103,and (ii) a light chain variable region comprising an amino acid sequenceselected from the group consisting of SEQ ID NO: 104 to 139.

In another embodiment, the humanized anti-Globo H antibody or anantigen-binding portion thereof comprises a heavy chain variable regioncomprising an amino acid sequence consisting of SEQ ID NO: 90, and alight chain variable region comprising an amino acid sequence consistingof SEQ ID NO: 135 (MK1 antibody).

Heavy Chain Variable Region of MK1 Series

(QVTLKESGPALVKPTQTLTLTCTFS************WIRQPPGKALEWLA*************LKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCAR*************WGQGTLVTVSS) (SEQ ID NO: 80)QVTLKESGPALVKPTQTLTLTCTFSGFSLSTFDMGVGWIRQPPGKALEWLAHIWWDDDKYYNPALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 81)QVTLKESGPALVKPTQTLTLTCTFSGFSLSTFDMGVGWIRQPPGKALEWLAHIWWDDDKYYNPALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARLSGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 82)QVTLKESGPALVKPTQTLTLTCTFSGFSLSTFDMGVGWIRQPPGKALEWLAHIWWDDDKYYNPALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLRSYYCDYWGQGTLVTVSS (SEQ ID NO: 83)QVTLKESGPALVKPTQTLTLTCTFSGSSLSTFDVGVGWIRQPPGKALEWLAHIWWDDDKYYNPALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 84)QVTLKESGPALVKPTQTLTLTCTFSGSSLSTFDVGVGWIRQPPGKALEWLAHIWWDDDKYYNPALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARLSGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 85)QVTLKESGPALVKPTQTLTLTCTFSGSSLSTFDVGVGWIRQPPGKALEWLAHIWWDDDKYYNPALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLRSYYCDYWGQGTLVTVSS (SEQ ID NO: 86)QVTLKESGPALVKPTQTLTLTCTFSGFSLGTFDLGIGWIRQPPGKALEWLAHIWWDDDKYYNPALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 87)QVTLKESGPALVKPTQTLTLTCTFSGFSLGTFDLGIGWIRQPPGKALEWLAHIWWDDDKYYNPALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARLSGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 88)QVTLKESGPALVKPTQTLTLTCTFSGFSLGTFDLGIGWIRQPPGKALEWLAHIWWDDDKYYNPALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLRSYYCDYWGQGTLVTVSS (SEQ ID NO: 89)QVTLKESGPALVKPTQTLTLTCTFSGFSLSTFDLGIGWIRQPPGKALEWLAHIWWDDDKYYNPALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLTSFYCDYWGQGTLVTVSS  (SEQ ID NO: 90)QVTLKESGPALVKPTQTLTLTCTFSGFSLSTFDLGIGWIRQPPGKALEWLAHIWWDDDKYYNPALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARLSGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 91)QVTLKESGPALVKPTQTLTLTCTFSGFSLSTFDLGIGWIRQPPGKALEWLAHIWWDDDKYYNPALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLRSYYCDYWGQGTLVTVSS (SEQ ID NO: 92)QVTLKESGPALVKPTQTLTLTCTFSGFSLSTFDMGVGWIRQPPGKALEWLAHIWGDDDKYYNPALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 93)QVTLKESGPALVKPTQTLTLTCTFSGFSLSTFDMGVGWIRQPPGKALEWLAHIWGDDDKYYNPALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARLSGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 94)QVTLKESGPALVKPTQTLTLTCTFSGFSLSTFDMGVGWIRQPPGKALEWLAHIWGDDDKYYNPALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLRSYYCDYWGQGTLVTVSS (SEQ ID NO: 95)QVTLKESGPALVKPTQTLTLTCTFSGSSLSTFDVGVGWIRQPPGKALEWLAHIWGDDDKYYNPALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLTSFYCDYWGQGTLVTVSS  (SEQ ID NO: 96)QVTLKESGPALVKPTQTLTLTCTFSGSSLSTFDVGVGWIRQPPGKALEWLAHIWGDDDKYYNPALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARLSGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 97)QVTLKESGPALVKPTQTLTLTCTFSGSSLSTFDVGVGWIRQPPGKALEWLAHIWGDDDKYYNPALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLRSYYCDYWGQGTLVTVSS (SEQ ID NO: 98)QVTLKESGPALVKPTQTLTLTCTFSGFSLGTFDLGIGWIRQPPGKALEWLAHIWGDDDKYYNPALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 99)QVTLKESGPALVKPTQTLTLTCTFSGFSLGTFDLGIGWIRQPPGKALEWLAHIWGDDDKYYNPALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARLSGNYLTSFYCDYWGQGTLVTVSS  (SEQ ID NO: 100)QVTLKESGPALVKPTQTLTLTCTFSGFSLGTFDLGIGWIRQPPGKALEWLAHIWGDDDKYYNPALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLRSYYCDYWGQGTLVTVSS  (SEQ ID NO: 101)QVTLKESGPALVKPTQTLTLTCTFSGFSLSTFDLGIGWIRQPPGKALEWLAHIWGDDDKYYNPALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 102)QVTLKESGPALVKPTQTLTLTCTFSGFSLSTFDLGIGWIRQPPGKALEWLAHIWGDDDKYYNPALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARLSGNYLTSFYCDYWGQGTLVTVSS (SEQ ID NO: 103)QVTLKESGPALVKPTQTLTLTCTFSGFSLSTFDLGIGWIRQPPGKALEWLAHIWGDDDKYYNPALKSRLTISKDTSKNQVVLTMTNMDPVDTATYYCARLYGNYLRSYYCDYWGQGTLVTVSS

Light Chain Variable Region of MK1 Series

(DVVMTQSPAFLSVTPGEKVTITC**********WYQQKPDQAPKLLIK*******GVPSRFSGSGSGTDFTFTISSLEAEDAATYYC*********FGQGTKLEIKR) (SEQ ID NO: 104)DVVMTQSPAFLSVTPGEKVTITCSASSSVSYMHWYQQKPDQAPKLLIKATSNLASGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQWSSNPFTFGQGTKLEIKR (SEQ ID NO: 105)DVVMTQSPAFLSVTPGEKVTITCSASSSVSYMHWYQQKPDQAPKLLIKATSNLASGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQWSSNPLTFGQGTKLEIKR (SEQ ID NO: 106)DVVMTQSPAFLSVTPGEKVTITCSASSSVSYMHWYQQKPDQAPKLLIKATSNLASGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQHLHIPYTFGQGTKLEIKR (SEQ ID NO: 107)DVVMTQSPAFLSVTPGEKVTITCSASSRVSYMHWYQQKPDQAPKLLIKATSNLASGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQWSSNPFTFGQGTKLEIKR (SEQ ID NO: 108)DVVMTQSPAFLSVTPGEKVTITCSASSRVSYMHWYQQKPDQAPKLLIKATSNLASGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQWSSNPLTFGQGTKLEIKR (SEQ ID NO: 109)DVVMTQSPAFLSVTPGEKVTITCSASSRVSYMHWYQQKPDQAPKLLIKATSNLASGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQHLHIPYTFGQGTKLEIKR (SEQ ID NO: 110)DVVMTQSPAFLSVTPGEKVTITCSARSSVSYMHWYQQKPDQAPKLLIKATSNLASGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQWSSNPFTFGQGTKLEIKR (SEQ ID NO: 111)DVVMTQSPAFLSVTPGEKVTITCSARSSVSYMHWYQQKPDQAPKLLIKATSNLASGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQWSSNPLTFGQGTKLEIKR (SEQ ID NO: 112)DVVMTQSPAFLSVTPGEKVTITCSARSSVSYMHWYQQKPDQAPKLLIKATSNLASGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQHLHIPYTFGQGTKLEIKR (SEQ ID NO: 113)DVVMTQSPAFLSVTPGEKVTITCRASSSVSYMHWYQQKPDQAPKLLIKATSNLASGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQWSSNPFTFGQGTKLEIKR (SEQ ID NO: 114)DVVMTQSPAFLSVTPGEKVTITCRASSSVSYMHWYQQKPDQAPKLLIKATSNLASGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQWSSNPLTFGQGTKLEIKR (SEQ ID NO: 115)DVVMTQSPAFLSVTPGEKVTITCRASSSVSYMHWYQQKPDQAPKLLIKATSNLASGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQHLHIPYTFGQGTKLEIKR (SEQ ID NO: 116)DVVMTQSPAFLSVTPGEKVTITCSASSSVSYMHWYQQKPDQAPKLLIKWTSDRYSGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQWSSNPFTFGQGTKLEIKR (SEQ ID NO: 117)DVVMTQSPAFLSVTPGEKVTITCSASSSVSYMHWYQQKPDQAPKLLIKWTSDRYSGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQWSSNPLTFGQGTKLEIKR (SEQ ID NO: 118)DVVMTQSPAFLSVTPGEKVTITCSASSSVSYMHWYQQKPDQAPKLLIKWTSDRYSGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQHLHIPYTFGQGTKLEIKR (SEQ ID NO: 119)DVVMTQSPAFLSVTPGEKVTITCSASSRVSYMHWYQQKPDQAPKLLIKWTSDRYSGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQWSSNPFTFGQGTKLEIKR (SEQ ID NO: 120)DVVMTQSPAFLSVTPGEKVTITCSASSRVSYMHWYQQKPDQAPKLLIKWTSDRYSGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQWSSNPLTFGQGTKLEIKR (SEQ ID NO: 121)DVVMTQSPAFLSVTPGEKVTITCSASSRVSYMHWYQQKPDQAPKLLIKWTSDRYSGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQHLHIPYTFGQGTKLEIKR (SEQ ID NO: 122)DVVMTQSPAFLSVTPGEKVTITCSARSSVSYMHWYQQKPDQAPKLLIKWTSDRYSGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQWSSNPFTFGQGTKLEIKR (SEQ ID NO: 123)DVVMTQSPAFLSVTPGEKVTITCSARSSVSYMHWYQQKPDQAPKLLIKWTSDRYSGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQWSSNPLTFGQGTKLEIKR (SEQ ID NO: 124)DVVMTQSPAFLSVTPGEKVTITCSARSSVSYMHWYQQKPDQAPKLLIKWTSDRYSGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQHLHIPYTFGQGTKLEIKR (SEQ ID NO: 125)DVVMTQSPAFLSVTPGEKVTITCRASSSVSYMHWYQQKPDQAPKLLIKWTSDRYSGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQWSSNPFTFGQGTKLEIKR (SEQ ID NO: 126)DVVMTQSPAFLSVTPGEKVTITCRASSSVSYMHWYQQKPDQAPKLLIKWTSDRYSGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQWSSNPLTFGQGTKLEIKR (SEQ ID NO: 127)DVVMTQSPAFLSVTPGEKVTITCRASSSVSYMHWYQQKPDQAPKLLIKWTSDRYSGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQHLHIPYTFGQGTKLEIKR (SEQ ID NO: 128)DVVMTQSPAFLSVTPGEKVTITCSASSSVSYMHWYQQKPDQAPKLLIKDTSKLASGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQWSSNPFTFGQGTKLEIKR (SEQ ID NO: 129)DVVMTQSPAFLSVTPGEKVTITCSASSSVSYMHWYQQKPDQAPKLLIKDTSKLASGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQWSSNPLTFGQGTKLEIKR (SEQ ID NO: 130)DVVMTQSPAFLSVTPGEKVTITCSASSSVSYMHWYQQKPDQAPKLLIKDTSKLASGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQHLHIPYTFGQGTKLEIKR (SEQ ID NO: 131)DVVMTQSPAFLSVTPGEKVTITCSASSRVSYMHWYQQKPDQAPKLLIKDTSKLASGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQWSSNPFTFGQGTKLEIKR (SEQ ID NO: 132)DVVMTQSPAFLSVTPGEKVTITCSASSRVSYMHWYQQKPDQAPKLLIKDTSKLASGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQWSSNPLTFGQGTKLEIKR (SEQ ID NO: 133)DVVMTQSPAFLSVTPGEKVTITCSASSRVSYMHWYQQKPDQAPKLLIKDTSKLASGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQHLHIPYTFGQGTKLEIKR (SEQ ID NO: 134)DVVMTQSPAFLSVTPGEKVTITCSARSSVSYMHWYQQKPDQAPKLLIKDTSKLASGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQWSSNPFTFGQGTKLEIKR (SEQ ID NO: 135)DVVMTQSPAFLSVTPGEKVTITCSARSSVSYMHWYQQKPDQAPKLLIKDTSKLASGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQWSSNPLTFGQGTKLEIKR (SEQ ID NO: 136)DVVMTQSPAFLSVTPGEKVTITCSARSSVSYMHWYQQKPDQAPKLLIKDTSKLASGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQHLHIPYTFGQGTKLEIKR (SEQ ID NO: 137)DVVMTQSPAFLSVTPGEKVTITCRASSSVSYMHWYQQKPDQAPKLLIKDTSKLASGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQWSSNPFTFGQGTKLEIKR (SEQ ID NO: 138)DVVMTQSPAFLSVTPGEKVTITCRASSSVSYMHWYQQKPDQAPKLLIKDTSKLASGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQWSSNPLTFGQGTKLEIKR (SEQ ID NO: 139)DVVMTQSPAFLSVTPGEKVTITCRASSSVSYMHWYQQKPDQAPKLLIKDTSKLASGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQHLHIPYTFGQGTKLEIKR

A humanized antibody has one or more amino acid residues from a sourcethat is non-human. The non-human amino acid residues are often referredto as “import” residues, and are typically taken from an “import”variable domain. Humanization can be performed generally followingconventional method known in the art, by substituting rodent CDRs or CDRsequences for the corresponding sequences of a human antibody.Accordingly, such “humanized” antibodies are antibodies whereinsubstantially less than an intact human variable domain has beensubstituted by the corresponding sequence from a non-human species. Inpractice, humanized antibodies are typically human antibodies in whichsome CDR residues and possibly some FR residues are substituted byresidues from analogous sites in non-human, for example, rodentantibodies.

The choice of human variable domains, both light and heavy, to be usedin making the humanized antibodies is very important to reduceantigenicity. The sequence of the variable domain of a rodent antibodyis screened against the entire library of known human variable-domainsequences. The human sequence which is closest to that of the rodent isthen accepted as the human framework (FR) for the humanized antibody.Another method uses a particular framework derived from the consensussequence of all human antibodies of a particular subgroup of light orheavy chains. The same framework may be used for several differenthumanized antibodies.

It is further important that antibodies be humanized with retention ofhigh affinity for the antigen and other favorable biological properties.To achieve this goal, according to a preferred method, humanizedantibodies are prepared by a process of analysis of the parentalsequences and various conceptual humanized products usingthree-dimensional models of the parental and humanized sequences.Three-dimensional immunoglobulin models are commonly available and arefamiliar to those skilled in the art. Computer programs are availablewhich illustrate and display probable three-dimensional conformationalstructures of selected candidate immunoglobulin sequences. Inspection ofthese displays permits analysis of the likely role of the residues inthe functioning of the candidate immunoglobulin sequence, i.e., theanalysis of residues that influence the ability of the candidateimmunoglobulin to bind its antigen. In this way, FR residues can beselected and combined from the recipient and import sequences so thatthe desired antibody characteristic, such as increased affinity for thetarget antigen(s), is achieved. In general, the CDR residues aredirectly and most substantially involved in influencing antigen binding.

In one embodiment, the present invention provides a chimeric anti-GloboH antibody or an antigen-binding portion thereof, comprising (i) a heavychain variable region comprising an amino acid sequence having at least85% identical to any of the amino acid sequences of SEQ ID NOs: 140-163wherein the last third sequence V of the amino acid sequences is changedas I, and (ii) a light chain variable region comprising an amino acidsequence having at least 80% identical to any of the amino acid sequenceof SEQ ID NO:164-199. Preferably, the sequence identity as mentionedabove is at least 90%, 91%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or99%.

In one embodiment, the present invention provides a chimeric anti-GloboH antibody, comprising (i) a heavy chain variable region comprising anamino acid sequence having at least 85% identical to the amino acidsequences of SEQ ID NO: 147 wherein the last third sequence V of theamino acid sequence is changed as I, and (ii) a light chain variableregion comprising an amino acid sequence having at least 85% identicalto the amino acid sequence of SEQ ID NO: 195. In a further embodiment,the present invention provides an isolated anti-Globo H antibody(cMZ-2), comprising (i) a heavy chain variable region comprising anamino acid sequence consisting of SEQ ID NO: 147 wherein the last thirdsequence V of the amino acid sequences is changed as I, and (ii) a lightchain variable region comprising an amino acid sequence consisting ofSEQ ID NO: 195.

The production of the chimeric antibody can be produced according toconventional methods known in the art. Methods for producing chimericantibodies are known in the art. See e.g., Morrison, Science 229:1202(1985); Oi et al., BioTechniques 4:214 (1986); Gillies et al., (1989) J.Immunol. Methods 125:191-202; U.S. Pat. Nos. 5,807,715; 4,816,567; and4,816,397. In addition, techniques developed for the production of“chimeric antibodies” (Morrison et al., 1984, Proc. Natl. Acad. Sci.81:851-855; Neuberger et al., 1984, Nature 312:604-608; Takeda et al.,1985, Nature 314:452-454 which are incorporated herein by reference intheir entireties) by splicing genes from a mouse antibody molecule ofappropriate antigen specificity together with genes from a humanantibody molecule of appropriate biological activity can be used. Insome embodiments, the constant domains of the light and heavy chains of,for example, a mouse monoclonal antibody can be substituted 1) for thoseregions of, for example, a human antibody to generate a chimericantibody or 2) for a non-immunoglobulin polypeptide to generate a fusionantibody. In other embodiments, the constant regions are truncated orremoved to generate the desired antibody fragment of a monoclonalantibody. Furthermore, site-directed or high-density mutagenesis of thevariable region can be used to optimize specificity, affinity, etc. of amonoclonal antibody.

Compositions of Antibodies of the Invention

In another aspect, the present invention provides a pharmaceuticalcomposition comprising an anti-Globo H antibody or an antigen-bindingportion thereof. The present antibody can also be formulated into apharmaceutical composition. In addition to the antibody or anantigen-binding portion thereof, the pharmaceutical composition furthercomprises a pharmaceutically acceptable carrier.

Administration of the anti-Globo H antibody or an antigen-bindingportion thereof described herein, can include formulation intopharmaceutical compositions or pharmaceutical formulations forparenteral administration, e.g., intravenous; mucosal, e.g., intranasal;ocular, or other mode of administration. In some embodiments, theanti-Globo H antibody or an antigen-binding portion thereof describedherein can be administered along with any pharmaceutically acceptablecarrier compound, material, or composition which results in an effectivetreatment in the subject. Thus, a pharmaceutical formulation/compositionfor use in the methods described herein can contain an anti-Globo Hantibody or an antigen-binding portion thereof as described herein incombination with one or more pharmaceutically acceptable carriers.

The phrase “pharmaceutically acceptable” refers to those compounds,materials, compositions, and/or dosage forms which are, within the scopeof sound medical judgment, suitable for use in contact with the tissuesof human beings and animals without excessive toxicity, irritation,allergic response, or other problem or complication, commensurate with areasonable benefit/risk ratio. The phrase “pharmaceutically acceptablecarrier” as used herein means a pharmaceutically acceptable material,composition or vehicle, such as a liquid or solid filler, diluent,excipient, solvent, media, encapsulating material, manufacturing aid(e.g., lubricant, talc magnesium, calcium or zinc stearate, or stericacid), or solvent encapsulating material, involved in maintaining thestability, solubility, or activity of, an anti-Globo H antibody or anantigen-binding portion thereof as described herein. Each carrier mustbe “acceptable” in the sense of being compatible with the otheringredients of the formulation and not injurious to the patient. Theterms “excipient”, “carrier”, “pharmaceutically acceptable carrier”, orthe like are used interchangeably herein.

The anti-Globo H antibody or an antigen-binding portion thereof asdescribed herein can be specially formulated for administration of thecompound to a subject in solid, liquid or gel form, including thoseadapted for the following: (1) parenteral administration, for example,by subcutaneous, intramuscular, intravenous or epidural injection as,for example, a sterile solution or suspension, or sustained-releaseformulation; (2) topical application, for example, as a cream, ointment,or a controlled-release patch or spray applied to the skin; (3)intravaginally or intrarectally, for example, as a pessary, cream orfoam; (4) ocularly; (5) transdermally; (6) transmucosally; or (79)nasally. Additionally, a anti-Globo H antibody or an antigen-bindingportion thereof as described herein can be implanted into a patient orinjected using a drug delivery system.

According to various working examples presented below, adult C57BL/6mice (weight 20-25 grams) administered with the present antibody twice aweek achieved reduced tumor size on 3 to 21 days after inoculation.Hence, in certain embodiments of the present disclosure, thetherapeutically effective amount of the antibody for mice could beexpressed as 0.8-100 mg/kg body weight. HED of the above-mentionedmurine effective amount is about 0.65-81.5 mg/kg body weight. Accordingto various embodiments of the present disclosure, when the subject ishuman, the therapeutically effective amount of the antibody can be atleast 1 mg/kg. Depending on the type and severity of the disease, about1 mg/kg to 150 mg/kg (e.g., 0.1-20 mg/kg) of an anti-Globo H antibody oran antigen-binding portion thereof as described herein is an initialcandidate dosage for administration to a subject, whether, for example,by one or more separate administrations, or by continuous infusion. Atypical daily dosage might range from about 1 mg/kg to about 100 mg/kgor more, depending on the factors mentioned above. Typical dosagesinclude, for example, 5 mg/kg, 10 mg/kg, 20 mg/kg, and 30 mg/kg. Forrepeated administrations over several days or longer, depending on thecondition, the treatment is sustained until, for example, the cancer istreated, as measured by the methods described above or known in the art.

Modes of Administration

In another aspect, the invention provides a method of treating and/orpreventing a cancer, comprising administering to a subject in needthereof a therapeutically effective amount of an anti-Globo H antibodyor an antigen-binding portion thereof as described herein or apharmaceutical composition comprising an anti-Globo H antibody or anantigen-binding portion thereof as described herein.

The administration of the antibody or the pharmaceutical compositioncomprising the same to a subject confers passive protection to thesubject and thereby provides the intended therapeutic effect to acancer, such as tumor-associated carbohydrate-expressing cancer;preferably, breast cancer, ovarian cancer, pancreatic cancer, prostatecancer, colorectal cancer and lung cancer.

In some embodiments, the anti-Globo H antibody or an antigen-bindingportion thereof as described herein or a pharmaceutical compositioncomprising an anti-Globo H antibody or an antigen-binding portionthereof as described herein is administered to a subject having a cancerto be inhibited by any mode of administration that delivers the agentsystemically or to a desired surface or target, and can include, but isnot limited to, injection, infusion, instillation, and inhalationadministration. To the extent that anti-Globo H antibody or anantigen-binding portion thereof as described herein or a pharmaceuticalcomposition comprising an anti-Globo H antibody or an antigen-bindingportion thereof as described herein can be protected from inactivationin the gut, oral administration forms are also contemplated. “Injection”includes, without limitation, intravenous, intramuscular, intraarterial,intrathecal, intraventricular, intracapsular, intraorbital,intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous,subcuticular, intraarticular, sub capsular, subarachnoid, intraspinal,intracerebro spinal, and intrasternal injection and infusion. Inpreferred embodiments, the anti-Globo H antibody or an antigen-bindingportion thereof as described herein or a pharmaceutical compositioncomprising an anti-Globo H antibody or an antigen-binding portionthereof as described herein for use in the methods described herein areadministered by intravenous infusion or injection.

The phrases “parenteral administration” and “administered parenterally”as used herein, refer to modes of administration other than enteral andtopical administration, usually by injection. The phrases “systemicadministration,” “administered systemically”, “peripheraladministration” and “administered peripherally” as used herein refer tothe administration of the bispecific or multispecific polypeptide agentother than directly into a target site, tissue, or organ, such as atumor site, such that it enters the subject's circulatory system and,thus, is subject to metabolism and other like processes.

The methods provided herein for inhibiting or treating cancer in subjecthaving or at risk for cancer by administering to the subject atherapeutically effective amount of an anti-Globo H antibody or anantigen-binding portion thereof as described herein or a pharmaceuticalcomposition comprising an anti-Globo H antibody or an antigen-bindingportion thereof as described herein, can, in some embodiments, furthercomprise administration one or more additional treatments such asangiogenic inhibitors, chemotherapy, radiation, surgery, or othertreatments known to those of skill in the art to prevent and/or treatcancer.

The following Examples are provided to elucidate certain aspects of thepresent invention and to aid those of skilled in the art in practicingthis invention. These Examples are in no way to be considered to limitthe scope of the invention in any manner. Without further elaboration,it is believed that one skilled in the art can, based on the descriptionherein, utilize the present invention to its fullest extent. Allpublications cited herein are hereby incorporated by reference in theirentirety.

EXAMPLE Example 1 Production of Monoclonal Antibody, Chimeric Antibodyand Humanized Antibody Against Globo H

Adult female C57BL/6 mice (n=3 each group; 5 weeks old; average weight16-20 grams; purchased from Biolasco, Taiwan) were immunized bysubcutaneous injection with 6 μg of Globo H-PADRE glycopeptide whereinPARDE represents a polypeptide of AKXVAAWTLKAAA (SEQ ID NO:238), and 50μl of complete Freund's adjuvant (CFA; from Sigma). Four immunizationswere given at a 2-week interval. Three days after the fourthimmunization, immunized splenocytes were harvest and washed withserum-free medium. Subsequently, 1×10⁸ of single cell suspendedsplenocytes were mixed with 2×10⁷ of FO cells, and cell fusion wasperformed in 1 ml of 50% PEG 1500 solution (Roche) at 37° C. followed bydrop-wise addition of 13 ml of warmed RPMI medium (Gibco). Fused cellswere centrifuged and washed twice with complete medium. Cells were thenre-suspended in complete medium with 1× BM-Conditioned H1 Hybridomacloning supplement (Roche) and seeded into 96-well plates. For targetspecific B cell-myeloma cells fusion, immunized splenocytes wereincubated with Globo H-biotin (10 μg/ml) in serum-free RPMI medium for 3hours at 4° C. After being washed three times with the same medium,Globo H-binotin-bearing cells were resuspended at a concentration of1×10⁸ cells/ml and incubated with streptavidin (50 μg/ml) for 30 minutesat 4° C. Meanwhile, FO cells were incubated with 50 μg/ml of NHS-biotinfor 1 hour at 4° C. Both treated cells were then washed three times withserum-free RPMI medium. Then, 1×10⁸ splenocytes and 2×10⁷ FO cells weremixed together, and chemical cell fusion was performed as describeabove. After cell fusion, cells were cultured in RPMI 1640 mediumcontaining 1× HAT medium (Gibco) for further selection.

Monoclonal antibody-producing hybridoma cell lines were screened throughlimited dilution by ELISA assay on plate coated with Globo H-biotinantigen. Five clones (named MZ-1 to MZ-5, respectively) capable ofsecreting high-titers of anti-Globo H IgG or IgM antibodies wereobtained.

Supernatants from these hybridoma lines were also subjected to cellbinding assay. Briefly, 100 μl of the supernatant from the hybridomaculture was incubated with 2×105 of MCF-7 cells and then analyzed byflow cytometry with appropriate fluorescent secondary antibody mentionedbelow. The cells were washed once with 2 ml of 1× PBS. Aftercentrifugation, the wash buffer was discarded and cells were resuspendedin 100 μl of 1:100 diluted PE anti-mouse IgG-Fc (Jackson immunoresearch)or 100 μl of 1:100 diluted PE anti-mouse IgM (eBioscience) and incubatedagain at room temperature for 20 minutes. The cells were washed with PBSand resuspended in 200 μl of 1× PBS after centrifugation. The binding ofantibodies with cells were detected by flow cytometry. The resultsprovided in FIGS. 1A to E reveal that the monoclonal antibody producedby MZ-2 hybridoma (see FIG. 1C) (hereinafter, the MZ-2 antibody) hadgood binding affinity to MCF-7 cells. For comparison purpose, acommercially available anti-Globo H IgG3 antibody (see FIG. E), VK9antibody (see FIG. B) (eBioscience), was also analyzed.

Variable regions of heavy and light chains of MZ-2 antibody were firstcloned to human IgG1 and kappa chain conserved region expression vectorsto form mouse-human chimeric MZ-2 (cMZ-2). The cMZ-2 antibody has beengone through humanization by CDR-grafting based on structure analysisand sequence homology. The humanized MZ-2 antibody (hMZ-2) could bereadily expressed as a recombinant IgG in mammalian cells and retainsthe antigen-binding affinity and specificity of the parent mouseantibody. We further improved its binding affinity with selectivemutations of CDR and framework sequences that flank the CDR. Thehumanized construct was further transfected into FO cells byelectroporation and selected by associated antibiotics to generatestable expression clones. Antibodies produced from the FO cells werechecked by flow cytometry analysis. To obtain large-scale hMZ-2antibodies for anti-tumor assay in vivo, 1×106 of hMZ-2 expression FOcells were first i.p. injected into NOD/SCID mice. Ascites from thesemice were harvest after 2 weeks and passed through protein G agarosecolumn to purify the generated antibody. The thus-obtained hMZ-2antibodies were sequenced. The hMZ-2 antibody, hMz-2Lw (variable regionof heavy chain: SEQ ID No. 27; variable region of light chain: SEQ IDNo. 75 and MK1 (variable region of heavy chain: SEQ ID No. 290; variableregion of light chain: SEQ ID No.135), were used in the followingexamples.

Example 2 Binding Affinity of MZ-2 Monoclonal Antibody to PrimaryOvarian Cancer Cells

Primary human ovarian cancer tissues were obtained under the approval ofIRB and patient consent. Ovarian cancer tissues were digested by MASCTumor Dissociation kit (MASC, 130-095-929). The single cell suspensionwas stained with MZ-2 monoclonal antibody (10 μg/ml; 100 μl) for 30minutes at 4° C., and stained with secondary antibody anti-mouse IgG1 PE(1:50; eBioscience, 2-405-82), then fixed with 4% paraformaldehyde for 1min at RT. The cell were then stained with FITC-conjugated mouseanti-human EpCAM (1:50; biolegend, 324204), and mouse anti-human SSEA4(1:50;biolegend, 330408) for 30 min at 4° C. Flow cytometric analysiswas conducted on a Becton Dickinson FACScan (BD FACSCalibur). FIG. 2demonstrate that the present MZ-2 monoclonal antibody exhibited goodbinding to primary ovarian cancer cells. Most epithelial primary cancercells were stained with either MZ-2 or SSEA4 antibody showing byrepresentative flow cytometry. EpCAM staining here represents a markerof epithelial cell.

Example 3 Specificity Assay of MZ-2 Antibody

Biotinylated-carbohydrates were conjugated on BD Cytometric Beads viaAvidin by Functional Bead Conjugation Buffer Set (BD). First, 75 μL ofthe functional beads were sonicated and incubated with 1.9 μL of 1M DTTat room temperature for 1 hr. At the same time, 20 uL of eachbiotin-carbohydrate (0.2 mg/mL) was mixed with 90 μg of maleimideactivated neutrAvidin (1 mg/mL in coupling buffer; Pierce) and incubatedat room temperature for 1 hr. The beads were washed 3 times with 1 mL ofcoupling buffer and resuspended in 20 μL of coupling buffer. Thecarbohydrate were then mixed with designed beads and incubated withfurther one hour. 2 μL of N-Ethylmaleimide (2 mg/mL on DMSO, Pierce) wasthen added into the conjugated beads and incubated for further 15 min.After 3 time wash, the beads were suspended in 500 μL of storage bufferand ready for use.

For detecting the specificity and relative titer of anti-Globo Hantibody, each set of carbohydrate-conjugated beads were mixed anddiluted to 1:50 of each. 50 μL of bead mixture was then transfer to Vbottom 96-well plate, and mixed with 50 μL of 1:1000 diluted serum or 1μg/mL of MZ-2 antibody. After incubation for 30 min, the beads werewashed with 150 μL of wash buffer and further stained with 100 μL ofPE-conjugated 2Ab (Jackson Immunoresearch). The binding of anti-Globo Hantibody were detected by FACS. FIG. 3 show the specificity of MZ-2antibody is indicated by the fact that MZ-2 and chimeric MZ-2 antibodiesonly bind to Globo H-conjugated beads, but not SSEA3- orSSEA4-conjugated beads.

Example 4 Simulation of Protein Folding in Mouse and Humanized MZ-2Monoclonal Antibody

The 3-D structure of variable regions in original and humanized MZ-2clones were simulated by Prediction of Immunoglobulin Structure onlineprogram. FIGS. 4A and B show the simulation of protein folding of mouseMZ-2 monoclonal antibody (FIG. 4A) and humanized MZ-2 monoclonalantibodies (FIG. 4B).

Example 5 Association and Dissociation Curve Fitting and KD Calculationof Antibodies of the Invention

The binding affinity of anti-Globo H antibody was detected by Biolayerinterferometry using FortrBio OcTet system. Briefly, the sensors werefirst soaked in 20 μM of biotinylated Globo H to coat Globo H on theirsurface. The MZ-2 clone (mMZ-2), mouse-human chimerical MZ-2 clone(cMZ-2), and humanized MZ-2 clones (hMZ-2L, MK-1 or hMZ-2Lw) wereserially diluted into 1333, 444.4, 148.1, 49.4 and 16.5 nM, andincubated with Globo H coated sensors separately. 10 mM of Glycine (pH1.5) was used as Regeneration buffer. FIGS. 5A and B show biacore fullbinding kinetic analysis of MZ-2 antibodies were carried out. Detailedbinding kinetic parameters (association rate, ka, dissociation rate, kd,and affinity constant, KD) were determined by full kinetic analysis.Sensograms for each antibody are shown. The KD values of three humanizedMZ-2 antibodies (hMZ-2L (see FIG. 5C), MK-1 (see FIG. 5D) and hMZ-2Lw(see FIG. 5E)) and chimeric MZ-2 (cMZ-2) antibody (see FIG. 5B) aresimilar.

Example 6 CDC Assay of Chimeric MZ-2 (IgG1 Kappa) on Breast MCF-7 Cells

MCF-7, TOV21G Globo H(+), and HPAC cells were adjusted to 2×106 cells/mLin RPMI 1640 medium and aliquot 50 μL of diluted cells into flow tubes.The chimeric MZ-2 antibody was diluted to 2× designed concentration inRPMI 1640 medium and aliquot 50 μL to each flow tube with cancer cells.Human IgG (Fitzgerald, 31-AI06) was diluted into the same concentrationas a control. After 15 min, 100 μL of normal human serum from healthyhuman donor was added into each tube and incubated at 37° C. for 2 hr.The cells were washed once with complete medium and resuspended in 200μL of complete medium with 0.5 μg/mL propidium iodide. The percentage ofdead cells was analyzed using FACS.

Results of complement-dependent cytotoxicity (CDC) assay, as reported inFIG. 6, indicate that the human serum containing more than 1.6 μg/ml ofhMZ-2Lw antibody elicited complement-dependent cytotoxicity in breastcancer, MCF-7 cells. The CDC increased in a dose-dependent manner.Similar trends were also observed in ovarian and pancreatic cancer cellsin which hMZ-2Lw antibody in a concentration higher than about 10 and 20μg/ml resulted in a dose-dependent cytotoxicity in human ovarian cancercell line TOV21G (FIG. 7), or the pancreatic cancer cell line, HPAC (seeFIG. 8).

Example 7 ADCC Assay of hMZ-2Lw and MK1 Antibody on MCF-7 Cells

Briefly, 7.5×10³ (100 uL) of each cells were pre-seeded into thedesigned wells of 96-well assay plate (Corning Cat. #3917) and incubatedovernight in a CO2 incubator at 37° C. The humanized anti-Globo Hantibody clone hMZ-2Lw or MK1 was diluted into 3× highest concentrationof assay and 4× serial dilution by 8 times. Noraml human IgG in the sameconcentration was used as control. ADCC assay was performed by ADCCReporter Bioassay kit (Promega Cat. #G7010) at a E/T ratio of 10:1 anddetected by chemoluminescent reader (EnSpire 2300, PerkinElmer). Thefolds of induced ADCC response were calculated as equation below:

Calculate Fold of Induction=RLU (induced-background)/RLU (no antibodycontrol-background)

FIGS. 9A and B and FIGS. 10A and B respectively summarizes theantigen-dependent cell-mediated cytotoxicity (ADCC) activity of thepresent hMZ-2Lw and MK1 antibodies on MCF-7 cells (breast cancer cellline) and TOV21G cells (ovarian cancer cell line). In general, theseresults demonstrate that the present humanized MZ-2 antibody may elicitefficient and dose-dependent ADCC in MCF-7 and TOV21G cells.

Example 8 Anti-Tumor Effect of Humanized MZ-2 Antibody, hMZ-2Lw and MK1

For establishing an intra-peritoneal ovarian tumor model, 1×10⁶ ofTOV21G cells were intraperitoneally (i.p.) injected into 5-week-oldfemale NU/NU mice (BioLASCO Taiwan). Two days later, mice were dividedinto 4 groups and administered with either 100 μg (at a therapeutic doseof 5 mg/kg) of Human IgG, anti-GloboH antibodies hMZ-2Lw or MK1 twice aweek through tail vein (i.v.) route. Untreated mice were set as control.To monitor the tumor growth, tumor bearing mice were i.p. injected 200μL of luciferin (3.9 mg/ml) and the chemoluminance intensity of eachmouse was detected by a non-invasive IVIS system (Xenogen) with fixedexposure condition in each different batch of experiment. Representativephotograph in FIG. 11 illustrates administration of both hMZ-2Lw and MK1antibodies significantly inhibit the tumor growth, while control IgGantibody did not substantially affect the tumor growth. However, thepresent hMZ-2 antibody significantly reduced the tumor size. By 24 daysafter treatment, tumor was nearly eliminated in mice given hMZ-2Lwantibody.

For establishing a subcutaneous breast tumor model, 17b-estradiol(Innovative Research of America, SE-121) was first subcutaneously (s.c.)implanted into NU/NU mice (BioLASCO Taiwan). Three days later, 3×10⁶MCF-7 cells mixed with matrigel were s.c. implanted (xenograft) intomice. Eighteen days after tumor challenge, mice were divided into 3groups (n=7), and treated by a therapeutic dose (5 mg/kg) of human IgG(100 μg/mouse) or 100 μg/mouse of hMZ-2Lw antibody i.v. (through tailvein) twice a week. Mice without treatment were set as control. Tumorsize was measured weekly by calipers. The results of hMZ-2 antibody inbreast cancer subcutaneous model MCF-7 cell are shown in FIG. 12. FIGS.13A and B shows the results of hMZ-2 antibody in pancreatic cancersubcutaneous model HPAC cell.

Example 9 MZ-2 Antibody Inhibits Migration of Globo H-Expressing TOV21GCells (I)

Briefly, 2×10⁵ of Globo H-positive TOV21G cells were mixed with 5 μg ofmouse IgG1 isotype or MZ-2 antibody (hMZ-2Lw) in 1 mL of culture mediumin and incubated at 37° C. for 15 min. These cells were then washedthree times with 1× PBS and resuspended to 2×10⁵ cells/mL in culturemedium. One hundred micro liters of cells were transferred into theinserts of transwell (Corning) and incubated in CO₂ incubator for 24 hr.Cells without antibody treatment, or Globo H-negative TOV21G cells wereused as controls. Those cells on the top side of transwell membrane wereremoved by cotton rod. Migrated cells through the membrane (bottom side)were fixed with 4% formaldehyde and stained with crystal violate. Thenumbers of migrated cells were counted by tissue scanner(TissueGnostics). FIG. 14 shows that MZ-2 antibody inhibits migration ofGlobo H-expressing TOV21G cells.

Example 10 MZ-2 Antibody Inhibits Migration of Globo H-Expressing TOV21GCells (II)

Briefly, 2×10⁵ of Globo H-expressing TOV21G cells were mixed with 5 μgof mouse IgG1 isotype (eBioscience) or MZ-2 antibody (hMZ-2Lw) in 0.5 mLof culture medium and incubated at 37° C. for 15 min. Those cells werethen washed three times with 1× PBS, and resuspended to 4×10⁵ cells/mLin culture medium. One hundred micro liters of cells were transferredinto a well of wound healing culture-inserts (Ibid Cat #80241) in a24-well plate and incubated for 8 hours in CO₂ incubator. Cells withoutantibody treatment, or Globo H-negative TOV21G cells were used ascontrols. The insert were then removed and the cells were kept culturedfor another 18 hours. Cell images were took by CCD camera from 0 hr to18 hr after insert removal. Note that the gap of cells treated by MZ-2antibody is larger than that treated by IgG1 isotype control. FIG. 15shows that MZ-2 antibody inhibits migration of Globo H-expressing TOV21Gcells.

Example 11 Assays of Other Antibodies of the Invention

The antibodies of the invention having the heavy chain variable regionsand the light chain variable regions as described herein were subjectedto the binding affinity assay of Example 2, association and dissociationassay of Example 5, CDC assay of Example 6, ADCC assay of Example 7 andanti-tumor assay of Example 8 and show similar results to cMZ-2antibody, hMZ-2Lw antibody and MK-1 antibody. For example, the KD valueof the antibody having a heavy chain variable region of SEQ ID NO: 27,and a light chain variable region of SEQ ID NO: 231 is in the range of1×10⁻⁷ to 1×10⁻¹⁰ nM.

1-26. (canceled)
 27. An anti-Globo H antibody or antigen-binding portionthereof comprising: (a) H-CDR1 comprising an amino acid sequenceselected from the group consisting of SEQ ID NO: 1, 2, 3, 4, and avariant of at least 90% identity to any one of SEQ ID NOs: 1-4; (b)H-CDR2 comprising an amino acid sequence selected from the groupconsisting of SEQ ID NO: 5, 6, and a variant of at least 90% identity toany one of SEQ ID NOs: 5-6; (c) H-CDR3 comprising an amino acid sequenceselected from the group consisting of SEQ ID NO: 7, 8, 9, and a variantof at least 90% identity to any one of SEQ ID NOs: 7-9; (d) L-CDR1comprising an amino acid sequence selected from the group consisting ofSEQ ID NO: 10, 11, 12, 13 and a variant of at least 90% identity to anyone of SEQ ID NOs: 10-13; (e) L-CDR2 comprising an amino acid sequenceselected from the group consisting of SEQ ID NO: 14, 15, 16, and avariant of at least 90% identity to any one of SEQ ID NOs: 14-16; and(f) L-CDR3 comprising an amino acid sequence selected from the groupconsisting of SEQ ID NO: 17, 18, 19, and a variant of at least 90%identity to any one of SEQ ID NOs: 17-19.
 28. The antibody orantigen-binding portion of claim 1 which is a monoclonal antibody, achimeric antibody, or a humanized antibody.
 29. The antibody orantigen-binding portion of claim 1 comprising (i) a heavy chain variableregion comprising H-CDR1 selected from the group consisting of SEQ IDNOs: 1-4, H-CDR2 selected from the group consisting of SEQ ID NOs: 5-6,and H-CDR3 selected from the group consisting of SEQ ID NOs: 7-9; and(ii) a light chain variable region comprising L-CDR1 selected from thegroup consisting of SEQ ID NOs: 10-13, L-CDR2 selected from the groupconsisting of SEQ ID NOs: 14-16, and L-CDR3 selected from the groupconsisting of SEQ ID NOs: 17-19.
 30. The antibody or antigen-bindingportion of claim 1, wherein H-CDR1 comprises SEQ ID NO: 3; H-CRD2comprises SEQ ID NO: 5; H-CDR3 comprises SEQ ID NO: 8; L-CDR1 comprisesSEQ ID NO:12; L-CDR2 comprises SEQ ID NO:16; and L-CDR3 comprises SEQ IDNO:
 18. 31. The antibody or antigen-binding portion of claim 1comprising (i) a heavy chain variable region comprising an amino acidsequence at least 85% identical to an amino acid sequence selected fromSEQ ID NOs: 20-43; and (ii) a light chain variable region comprising anamino acid sequence at least 80% identical to an amino acid sequenceselected from SEQ ID NO: 44-79, and 200-235.
 32. The antibody orantigen-binding portion of claim 31 comprising (i) a heavy chainvariable region comprising an amino acid sequence consisting of SEQ IDNO: 27, and a light chain variable region comprising an amino acidsequence consisting of SEQ ID NO: 75; or (ii) a heavy chain variableregion comprising an amino acid sequence consisting of SEQ ID NO: 27,and a light chain variable region comprising an amino acid sequenceconsisting of SEQ ID NO:
 231. 33. The antibody or antigen-bindingportion of claim 1 comprising (i) a heavy chain variable regioncomprising an amino acid sequence at least 85% identical to an aminoacid sequence selected from SEQ ID NOs: 140-163; and (ii) a light chainvariable region comprising an amino acid sequence at least 80% identicalto an amino acid sequence selected from SEQ ID NO: 164-199.
 34. Theantibody or antigen-binding portion of claim 33 comprising (i) a heavychain variable region comprising an amino acid sequence having at least85% identical to the amino acid sequences of SEQ ID NO: 147, and (ii) alight chain variable region comprising an amino acid sequence having atleast 80% identical to the amino acid sequence of SEQ ID NO:
 195. 35.The antibody or antigen-binding portion of claim 33 comprising (i) aheavy chain variable region comprising an amino acid sequence consistingof SEQ ID NO: 147; and (ii) a light chain variable region comprising anamino acid sequence consisting of SEQ ID NO:
 195. 36. The antibody orantigen-binding portion of claim 1 comprising (i) a heavy chain variableregion comprising an amino acid sequence at least 85% identical to anamino acid sequence selected from SEQ ID NOs: 80-103; and (ii) a lightchain variable region comprising an amino acid sequence at least 85%identical to an amino acid sequence of SEQ ID NOs: 104-139.
 37. Theantibody or antigen-binding portion of claim 36 comprising (i) a heavychain variable region comprising an amino acid sequence selected fromSEQ ID NOs: 80-103; and (ii) a light chain variable region comprising anamino acid sequence selected from SEQ ID NOs: 104-139.
 38. The antibodyor antigen-binding portion of claim 37 comprising (i) a heavy chainvariable region comprising an amino acid sequence consisting of SEQ IDNO: 90; and (ii) a light chain variable region comprising an amino acidsequence consisting of SEQ ID NO:
 135. 39. A pharmaceutical compositioncomprising an antibody or antigen-binding portion of claim 1 and apharmaceutically acceptable carrier.
 40. The pharmaceutical compositionof claim 39, further comprising an anti-tumor drug.
 41. A method oftreating and/or preventing a cancer, comprising administering to asubject in need thereof a therapeutically effective amount of anantibody or antigen-binding portion of claim
 1. 42. The method of claim41, wherein the cancer is a tumor-associated carbohydrate-expressingcancer.
 43. The method of claim 42, wherein the cancer is ovariancancer, breast cancer, pancreatic cancer, prostate cancer, colorectalcancer, or lung cancer.
 44. The method of claim 41, wherein the canceris a migration cancer.
 45. The method of claim 41, further comprisingadministration of one or more additional treatments selected fromangiogenic inhibitor treatment, chemotherapy, radiation, and surgery.